Idh1 inhibitors for the treatment of haematological malgnancies and solid tumors

ABSTRACT

Provided are methods and compositions for treating cancers in patients carrying an IDH1 mutation or IDH2 mutation.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a national stage application under 35 U.S.C. 371 ofInternational Application No. PCT/US2016/064845, filed Dec. 2, 2018,which claims the benefit of the priority of U.S. Provisional ApplicationNo. 62/300,673, filed Feb. 26, 2016, the disclosure of each of which isincorporated herein by reference in its entirety.

FIELD

Provided herein are methods of treating malignancies includinghematological malignancies and solid tumors characterized by thepresence of a mutant allele of IDH1 or IDH2, and the absence of a FLT3mutation. In one embodiment, the methods for treating a malignancycomprise administering an IDH1 inhibitor or an IDH2 inhibitor incombination with one or more compounds that target a FLT3 pathway,wherein the malignancy is characterized by the presence of a mutantallele of IDH1 or IDH2 respectively, and a mutant FLT3.

In one aspect, provided herein is an IDH1 inhibitor for use in methodsof treating malignancies including hematological malignancies and solidtumors characterized by the presence of a mutant allele of IDH1, and theabsence of a FLT3 mutation. In one embodiment, an IDH1 inhibitor can beprovided in combination with one or more compounds that target a FLT3pathway for use in methods for treating a malignancy, wherein themalignancy is characterized by the presence of a mutant allele of IDH1,and a mutant FLT3.

BACKGROUND

Isocitrate dehydrogenases (IDHs) catalyze the oxidative decarboxylationof isocitrate to 2-oxoglutarate (i.e., α-ketoglutarate). These enzymesbelong to two distinct subclasses, one of which utilizes NAD(+) as theelectron acceptor and the other NADP(+). Five isocitrate dehydrogenaseshave been reported: three NAD(+)-dependent isocitrate dehydrogenases,which localize to the mitochondrial matrix, and two NADP(+)-dependentisocitrate dehydrogenases, one of which is mitochondrial and the otherpredominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer.

IDH1 (isocitrate dehydrogenase 1 (NADP+), cytosolic) is also known asIDH; IDP; IDCD; IDPC or PICD. The protein encoded by this gene is theNADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm andperoxisomes. It contains the PTS-1 peroxisomal targeting signalsequence. The presence of this enzyme in peroxisomes suggests roles inthe regeneration of NADPH for intraperoxisomal reductions, such as theconversion of 2, 4-dienoyl-CoAs to 3-enoyl-CoAs, as well as inperoxisomal reactions that consume 2-oxoglutarate, namely thealpha-hydroxylation of phytanic acid. The cytoplasmic enzyme serves asignificant role in cytoplasmic NADPH production.

The human IDH1 gene encodes a protein of 414 amino acids. The nucleotideand amino acid sequences for human IDH1 can be found as GenBank entriesNM_005896.2 and NP_005887.2 respectively. The nucleotide and amino acidsequences for IDH1 are also described in, e.g., Nekrutenko et al., Mol.Biol. Evol. 15:1674-1684(1998); Geisbrecht et al., J. Biol. Chem.274:30527-30533(1999); Wiemann et al., Genome Res. 11:422-435(2001); TheMGC Project Team, Genome Res. 14:2121-2127(2004); Lubec et al.,Submitted (December-2008) to UniProtKB; Kullmann et al., Submitted(June-1996) to the EMBL/GenBank/DDBJ databases; and Sjoeblom et al.,Science 314:268-274(2006).

IDH2 (isocitrate dehydrogenase 2 (NADP+), mitochondrial) is also knownas IDH; IDP; IDHM; IDPM; ICD-M; or mNADP-IDH. The protein encoded bythis gene is the NADP(+)-dependent isocitrate dehydrogenase found in themitochondria. It plays a role in intermediary metabolism and energyproduction. This protein may tightly associate or interact with thepyruvate dehydrogenase complex. Human IDH2 gene encodes a protein of 452amino acids. The nucleotide and amino acid sequences for IDH2 can befound as GenBank entries NM_002168.2 and NP_002159.2 respectively. Thenucleotide and amino acid sequence for human IDH2 are also described in,e.g., Huh et al., Submitted (November-1992) to the EMBL/GenBank/DDBJdatabases; and The MGC Project Team, Genome Res. 14:2121-2127 (2004).

Non-mutant, e.g., wild type, IDH1 and IDH2 catalyze the oxidativedecarboxylation of isocitrate to α-ketoglutarate (α-KG) thereby reducingNAD⁺ (NADP⁺) to NADH (NADPH), e.g., in the forward reaction:

Isocitrate+NAD⁺(NADP⁺)→α-KG+CO₂+NADH(NADPH)+H⁺.

It has been discovered that mutations of IDH1 and IDH2 present incertain cancer cells result in a new ability of the enzyme to catalyzethe NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate (2HG). 2HG is not formed by wild-type IDH1 orIDH2. The production of 2HG is believed to contribute to the formationand progression of cancer (Dang, L. et al., Nature 462:739-44, 2009).

The development of selective inhibitors of IDH1 or IDH2 mutant enzymeshas provided the possibility of therapeutic benefit to cancer patientscarrying the IDH1 or IDH2 mutations. There is a need for improvedtherapies for treating cancer patients carrying IDH1 or IDH2 mutations.

SUMMARY

In one embodiment, provided herein are methods of treating hematologicmalignancies by administering to a subject a therapeutically effectiveamount of a mutant IDH1 inhibitor, wherein the hematologic malignancy ischaracterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation. In one embodiment, the hematologic malignancy is anadvanced hematologic malignancy. In one embodiment, the hematologicmalignancy is acute myeloid leukemia (AML). In one embodiment, the AMLis relapsed or refractory. Also provided herein is a mutant IDH1inhibitor for use in the methods of treating hematologic malignancies,wherein the hematologic malignancy is characterized by the presence of amutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided herein are methods of treating hematologicmalignancies by administering to a subject a therapeutically effectiveamount of a mutant IDH2 inhibitor, wherein the hematologic malignancy ischaracterized by the presence of a mutant allele of IDH2 and the absenceof a FLT3 mutation. In one embodiment, the hematologic malignancy is anadvanced hematologic malignancy. In one embodiment, the hematologicmalignancy is AML.

In one embodiment, provided herein are methods of treating hematologicmalignancies by administering to a subject a therapeutically effectiveamount of a mutant IDH1 inhibitor in combination with a therapeuticallyeffective amount of one or more compounds that target a FLT3 pathway,wherein the hematologic malignancy is characterized by the presence of amutant allele of IDH1 and a mutant FLT3, for example an internal tandemduplication mutation in the JM domain-coding sequence of the FLT3 gene(FLT3-ITD) or a missense point mutation at the D835 residue and pointmutations, deletions and insertions in the codons surrounding D835within a TK domain of FLT3 (FLT3-KDM) and/or other missense mutations.In one embodiment, the hematologic malignancy is AML. Also providedherein is an IDH1 inhibitor in combination with a therapeuticallyeffective amount of one or more compounds that target a FLT3 pathway foruse in the methods of treating hematologic malignancies, wherein thehematologic malignancy is characterized by the presence of a mutantallele of IDH1 and a mutant FLT3.

In one embodiment, provided herein are methods of treating hematologicmalignancies by administering to a subject a therapeutically effectiveamount of a mutant IDH2 inhibitor in combination with a therapeuticallyeffective amount of one or more compounds that target a FLT3 pathway,wherein the hematologic malignancy is characterized by the presence of amutant allele of IDH2 and a mutant FLT3, for example FLT3-ITD orFLT3-KDM. In one embodiment, the hematologic malignancy is an advancedhematologic malignancy. In one embodiment, the hematologic malignancy isAML.

In one embodiment, provided herein is a method of treating hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), chronic myelomonocytic leukemia (CMML), myeloid sarcoma,multiple myeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, each characterized by the presence of a mutantallele of IDH2 and the absence of a FLT3 mutation, comprisingadministering to a subject a therapeutically effective amount of2-methyl-1-[(4-[6-(trifluoromethyl)pyridin-2-yl]-6-{[2-(trifluoromethyl)pyridin-4-yl]amino}-1,3,5-triazin-2-yl)amino]propan-2-ol,or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug or a polymorph thereof (COMPOUND 1). In oneembodiment, the hematologic malignancy is an advanced hematologicmalignancy. In one embodiment, the hematologic malignancy is AML.

In one embodiment, provided herein is a method of treating hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), myeloproliferative neoplasms (MPN), chronicmyelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias (B-ALL),or lymphoma (e.g., T-cell lymphoma), each characterized by the presenceof a mutant allele of IDH1 and the absence of a FLT3 mutation,comprising administering to a subject a therapeutically effective amountof(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide,or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug or a polymorph thereof (COMPOUND 2). In oneembodiment, the hematologic malignancy is an advanced hematologicmalignancy. In one embodiment, the hematologic malignancy is AML. In oneembodiment, the AML is relapsed or refractory.

Also provided herein is COMPOUND 2 for use in the method of treatinghematologic malignancies, such as acute myelogenous leukemia (AML),myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), myeloid sarcoma, multiplemyeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, each characterized by the presence of a mutantallele of IDH1 and the absence of a FLT3 mutation. In one embodiment,the advanced hematologic malignancy is AML, characterized by thepresence of a mutant allele of IDH1 and the absence of a FLT3 mutation.In one embodiment, the advanced hematologic malignancy is relapsed orrefractory AML, characterized by the presence of a mutant allele of IDH1and the absence of a FLT3 mutation.

In one aspect, provided is COMPOUND 2 for use in a method of treatingacute myelogenous leukemia (AML) characterized by the presence of amutant allele of IDH1 and the absence of a FLT3 mutation. In one aspect,provided is COMPOUND 2 for use in a method of treating relapsed orrefractory acute myelogenous leukemia (AML) characterized by thepresence of a mutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided is COMPOUND 2 for use in a method oftreating myelodysplastic syndrome (MDS) characterized by the presence ofa mutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided is COMPOUND 2 for use in a method oftreating chronic myelomonocytic leukemia (CMML) characterized by thepresence of a mutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided is COMPOUND 2 for use in a method oftreating myeloid sarcoma characterized by the presence of a mutantallele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided is COMPOUND 2 for use in a method oftreating multiple myeloma characterized by the presence of a mutantallele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided is COMPOUND 2 for use in a method oftreating lymphoma (e.g., T-cell lymphoma or B-cell lymphoma)characterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation

In one embodiment, provided is COMPOUND 2 for use in a method oftreating angioimmunoblastic T-cell lymphoma (AITL) characterized by thepresence of a mutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided is COMPOUND 2 for use in a method oftreating blastic plasmacytoid dendritic cell neoplasm characterized bythe presence of a mutant allele of IDH1 and the absence of a FLT3mutation.

In one embodiment, provided herein is a method of treating hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), chronic myelomonocytic leukemia (CMML), myeloid sarcoma,multiple myeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, each characterized by the presence of a mutantallele of IDH2 and a mutant FLT3, for example FLT3-ITD, comprisingadministering to the subject a therapeutically effective amount of2-methyl-1-[(4-[6-(trifluoromethyl)pyridin-2-yl]-6-{[2-(trifluoromethyl)pyridin-4-yl]amino}-1,3,5-triazin-2-yl)amino]propan-2-ol,or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug or a polymorph thereof (COMPOUND 1) in combinationwith a therapeutically effective amount of one or more compounds thattarget a FLT3 pathway. In one embodiment, COMPOUND 1 is administered tothe subject in combination with a therapeutically effective amount of aFLT3 inhibitor selected from quizartinib (AC220), sunitinib (SU11248),sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib (CEP-701),crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534),ASP2215, KW-2449, famitinib and DCC-2036. In one embodiment, thehematologic malignancy is an advanced hematologic malignancy. In oneembodiment, the hematologic malignancy is AML.

In one embodiment, provided herein is a method of treating hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS, myeloproliferative neoplasms (MPN), chronicmyelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias (B-ALL),or lymphoma (e.g., T-cell lymphoma), each characterized by the presenceof a mutant allele of IDH1 and a mutant FLT3, for example FLT3-ITD, in asubject comprising administering to the subject a therapeuticallyeffective amount of(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide,or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug or a polymorph thereof (COMPOUND 2) in combinationwith a therapeutically effective amount of one or more compounds thattarget a FLT3 pathway.

In one embodiment, provided herein is COMPOUND 2 in combination with oneor more compounds that target a FLT3 pathway for use in a method oftreating hematologic malignancies, such as acute myelogenous leukemia(AML), myelodysplastic syndrome (MDS), chronic myelomonocytic leukemia(CMML), myeloid sarcoma, multiple myeloma, lymphoma (e.g., T-celllymphoma or B-cell lymphoma), angioimmunoblastic T-cell lymphoma (AITL)or blastic plasmacytoid dendritic cell neoplasm, each characterized bythe presence of a mutant allele of IDH1 and a mutant FLT3. In oneembodiment, provided herein is COMPOUND 2 in combination with one ormore compounds that target a FLT3 pathway for use in a method oftreating acute myelogenous leukemia (AML) characterized by the presenceof a mutant allele of IDH1 and a mutant FLT3. In one embodiment,provided herein is COMPOUND 2 in combination with one or more compoundsthat target a FLT3 pathway for use in a method of treatingmyelodysplastic syndrome (MDS) characterized by the presence of a mutantallele of IDH1 and a mutant FLT3. In one embodiment, provided herein isa COMPOUND 2 in combination with one or more compounds that target aFLT3 pathway for use in the method of treating chronic myelomonocyticleukemia (CMML) characterized by the presence of a mutant allele of IDH1and a mutant FLT3. In certain embodiments, provided herein is COMPOUND 2in combination with one or more compounds that target a FLT3 pathway foruse in a method of treating myeloid sarcoma characterized by thepresence of a mutant allele of IDH1 and a mutant FLT3. In oneembodiment, provided herein is COMPOUND 2 in combination with one ormore compounds that target a FLT3 pathway for use in a method oftreating multiple myeloma characterized by the presence of a mutantallele of IDH1 and a mutant FLT3. In one embodiment, provided herein isCOMPOUND 2 in combination with one or more compounds that target a FLT3pathway for use in a method of treating lymphoma (e.g., T-cell lymphomaor B-cell lymphoma) characterized by the presence of a mutant allele ofIDH1 and a mutant FLT3. In one embodiment, provided herein is COMPOUND 2in combination with one or more compounds that target a FLT3 pathway foruse in a method of treating angioimmunoblastic T-cell lymphoma (AITL)characterized by the presence of a mutant allele of IDH1 and a mutantFLT3. In one embodiment, provided herein is COMPOUND 2 in combinationwith one or more compounds that target a FLT3 pathway for use in amethod of treating blastic plasmacytoid dendritic cell neoplasmcharacterized by the presence of a mutant allele of IDH1 and a mutantFLT3.

In one embodiment, COMPOUND 2 is administered to the subject incombination with a therapeutically effective amount of a FLT3 inhibitorselected from quizartinib (AC220), sunitinib (SU11248), sorafenib (BAY43-9006), midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib(CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215,KW-2449, famitinib and DCC-2036. In one embodiment, the hematologicmalignancy is an advanced hematologic malignancy. In one embodiment, thehematologic malignancy is AML. In an embodiment, the AML ischaracterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation. In one embodiment, the advanced hematologicmalignancy is relapsed or refractory AML, characterized by the presenceof a mutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, COMPOUND 2 is administered to the subject incombination with quizartinib (AC220). In one embodiment, COMPOUND 2 isadministered to the subject in combination with sunitinib (SU11248). Inone embodiment, COMPOUND 2 is administered to the subject in combinationwith sorafenib (BAY 43-9006). In one embodiment, COMPOUND 2 isadministered to the subject in combination with midostaurin (PKC412). Inone embodiment, COMPOUND 2 is administered to the subject in combinationwith lestaurtinib (CEP-701). In one embodiment, COMPOUND 2 isadministered to the subject in combination with crenolanib (CP-868596).In one embodiment, COMPOUND 2 is administered to the subject incombination with PLX3397. In one embodiment, COMPOUND 2 is administeredto the subject in combination with PLX3397. In one embodiment, COMPOUND2 is administered to the subject in combination with E6201. In oneembodiment, COMPOUND 2 is administered to the subject in combinationwith AKN-028. In one embodiment, COMPOUND 2 is administered to thesubject in combination with ponatinib (AP24534). In one embodiment,COMPOUND 2 is administered to the subject in combination with ASP2215.In one embodiment, COMPOUND 2 is administered to the subject incombination with KW-2449. In one embodiment, COMPOUND 2 is administeredto the subject in combination with famitinib. In one embodiment,COMPOUND 2 is administered to the subject in combination with DCC-2036.

In one embodiment, provided herein are methods of solid tumors byadministering to a subject a therapeutically effective amount of amutant IDH1 inhibitor, wherein the solid tumor is characterized by thepresence of a mutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, provided herein is a mutant IDH1 inhibitor for use ina method of treating solid tumors, wherein the solid tumor ischaracterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation, for example FLT3-ITD.

In one embodiment, provided herein are methods of treating solid tumorsby administering to a subject a therapeutically effective amount of amutant IDH2 inhibitor, wherein the solid tumor is characterized by thepresence of a mutant allele of IDH2 and the absence of a FLT3 mutation.In one embodiment, the solid tumor is an advanced solid tumor.

In one embodiment, provided herein are methods of treating solid tumorsby administering to a subject a therapeutically effective amount of amutant IDH1 inhibitor in combination with a therapeutically effectiveamount of one or more compounds that target a FLT3 pathway, wherein thesolid tumor is characterized by the presence of a mutant allele of IDH1and a mutant FLT3, for example FLT3-ITD. In one aspect, provided hereinis a mutant IDH1 inhibitor in combination with one or more compoundsthat target a FLT3 pathway for use in the methods of treating solidtumors, wherein the solid tumor is characterized by the presence of amutant IDH1 and a mutant FLT3, for example FLT3-ITD.

In one embodiment, provided herein are methods of treating solid tumorsby administering to a subject a therapeutically effective amount of amutant IDH2 inhibitor in combination with a therapeutically effectiveamount of one or more compounds that target a FLT3 pathway, wherein thesolid tumor is characterized by the presence of a mutant IDH2 and amutant FLT3, for example FLT3-ITD. In one embodiment, the solid tumor isan advanced solid tumor.

In one embodiment, provided herein is a method of treating solid tumors,such as glioma, melanoma, chondrosarcoma, or cholangiocarcinoma(e.g.,glioma), or treating angioimmunoblastic T-cell lymphoma (AITL), eachcharacterized by the presence of a mutant allele of IDH2 and the absenceof a FLT3 mutation, comprising administering to a subject atherapeutically effective amount of COMPOUND 1.

In one embodiment, provided herein is a method of treating solid tumors,such as glioma, melanoma, chondrosarcoma, cholangiocarcinoma (includingintrahepatic cholangiocarcinoma (IHCC), prostate cancer, colon cancer,or non-small cell lung cancer (NSCLC), each characterized by thepresence of a mutant allele of IDH1 and the absence of a FLT3 mutation,comprising administering to a subject a therapeutically effective amountof COMPOUND 2.

In one embodiment, provided herein is a method of treating solid tumors,such as glioma, melanoma, chondrosarcoma, or cholangiocarcinoma (e.g.,glioma), or treating angioimmunoblastic T-cell lymphoma (AITL), eachcharacterized by the presence of a mutant allele of IDH2 and a mutantFLT3, in a subject comprising administering to a subject atherapeutically effective amount of COMPOUND 1 in combination with atherapeutically effective amount of one or more compounds that target aFLT3 pathway. In one embodiment, COMPOUND 1 is administered to thesubject in combination with a therapeutically effective amount of a FLT3inhibitor selected from quizartinib (AC220), sunitinib (SU11248),sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib (CEP-701),crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534),ASP2215, KW-2449, famitinib and DCC-2036.

In one embodiment, provided herein is a method of treating solid tumors,such as glioma, melanoma, chondrosarcoma, cholangiocarcinoma (includingintrahepatic cholangiocarcinoma (IHCC), prostate cancer, colon cancer,or non-small cell lung cancer (NSCLC), each characterized by thepresence of a mutant allele of IDH1 and a mutant FLT3, for exampleFLT3-ITD, in a subject comprising administering to a subject atherapeutically effective amount of COMPOUND 2 in combination with atherapeutically effective amount of one or more compounds that target aFLT3 pathway. In one embodiment, COMPOUND 2 is administered to thesubject in combination with a therapeutically effective amount of a FLT3inhibitor selected from quizartinib (AC220), sunitinib (SU11248),sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib (CEP-701),crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534),ASP2215, KW-2449, famitinib and DCC-2036.

In one embodiment, COMPOUND 2 is administered to the subject incombination with quizartinib (AC220). In one embodiment, COMPOUND 2 isadministered to the subject in combination with sunitinib (SU11248). Inone embodiment, COMPOUND 2 is administered to the subject in combinationwith sorafenib (BAY 43-9006). In one embodiment, COMPOUND 2 isadministered to the subject in combination with midostaurin (PKC412). Inone embodiment, COMPOUND 2 is administered to the subject in combinationwith lestaurtinib (CEP-701). In one embodiment, COMPOUND 2 isadministered to the subject in combination with crenolanib (CP-868596).In one embodiment, COMPOUND 2 is administered to the subject incombination with PLX3397. In one embodiment, COMPOUND 2 is administeredto the subject in combination with E6201. In one embodiment, COMPOUND 2is administered to the subject in combination with AKN-028. In oneembodiment, COMPOUND 2 is administered to the subject in combinationwith ponatinib (AP24534). In one embodiment, COMPOUND 2 is administeredto the subject in combination with ASP2215. In one embodiment, COMPOUND2 is administered to the subject in combination with KW-2449. In oneembodiment, COMPOUND 2 is administered to the subject in combinationwith famitinib. In one embodiment, COMPOUND 2 is administered to thesubject in combination with DCC-2036.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an X-ray powder diffractogram (XPRD) of COMPOUND 1 form 1.

FIG. 2 is an X-ray powder diffractogram (XPRD) of COMPOUND 1 form 2.

FIG. 3 is an X-ray powder diffractogram (XPRD) of COMPOUND 1 form 3.

FIG. 4 is an X-ray powder diffractogram (XPRD) of COMPOUND 1 form 4.

FIG. 5 is an X-ray powder diffractogram (XPRD) of COMPOUND 1 form 5.

FIG. 6 is an X-ray powder diffractogram (XPRD) of COMPOUND 1 form 6.

FIG. 7 is an X-ray powder diffractogram (XPRD) of COMPOUND 2 form I.

FIG. 8 is a differential scanning calorimetry (DSC) profile of COMPOUND2 form I.

FIG. 9 is a thermal gravimetric analysis (TGA) profile of COMPOUND 2form I.

FIG. 10 is an X-ray powder diffractogram (XPRD) of COMPOUND 2 form II.

FIG. 11 is a differential scanning calorimetry (DSC) profile of COMPOUND2 form II.

FIG. 12 is a thermal gravimetric analysis (TGA) profile of COMPOUND 2form II.

FIG. 13 illustrates the comutations, including FLT3 mutations, insamples treated with COMPOUND 1 according to response categories.

FIG. 14 illustrates the comutations, including FLT3 mutations in samplestreated with COMPOUND 2 according to response categories.

DETAILED DESCRIPTION

The details of construction and the arrangement of components set forthin the following description or illustrated in the drawings are notmeant to be limiting. Other embodiments and different ways to practicethe invention are expressly included. Also, the phraseology andterminology used herein is for the purpose of description and should notbe regarded as limiting. The use of “including,” “comprising,” or“having,” “containing”, “involving”, and variations thereof herein, ismeant to encompass the items listed thereafter and equivalents thereofas well as additional items.

Definitions

The term a “mutant IDH2 inhibitor” or “inhibitor of IDH2 mutant(s)”means a molecule e.g., a polypeptide, peptide, or small molecule (e.g.,a molecule of less than 1,000 daltons), or aptomer, that binds to anIDH2 mutant subunit and inhibits neoactivity, e.g., by inhibitingformation of a dimer, e.g., a homodimer of mutant IDH2 subunits or aheterodimer of a mutant and a wildype subunit. In some embodiments, theneoactivity inhibition is at least about 60%, 70%, 80%, 90%, 95% or 99%as compared to the activity in the absence of the mutant IDH2 inhibitor.In one embodiment, the mutant IDH2 inhibitor is COMPOUND 1.

The term a “mutant IDH1 inhibitor” or “inhibitor of IDH1 mutant(s)”means a molecule e.g., a polypeptide, peptide, or small molecule (e.g.,a molecule of less than 1,000 daltons), or aptomer, that binds to anIDH1 mutant subunit and inhibits neoactivity, e.g., by inhibitingformation of a dimer, e.g., a homodimer of mutant IDH1 subunits or aheterodimer of a mutant and a wildype subunit. In some embodiments, theneoactivity inhibition is at least about 60%, 70%, 80%, 90%, 95% or 99%as compared to the activity in the absence of the mutant IDH1 inhibitor.In one embodiment, the mutant IDH1 inhibitor is COMPOUND 2.

As used herein, the term “wild type” refers to the typical or mostcommon form of a characteristic (for example, gene sequence or presence,or protein sequence, presence, level or activity), as it occurs innature, and the reference against which all others are compared. As willbe understood by one skilled in the art, when used herein, wild typerefers to the typical gene sequence(s) or gene expression levels as theymost commonly occur in nature.

As used herein, “co-occurring mutation” refers to one or more genemutations that are present in a cancer subject herein in addition to anIDH1 or an IDH2 mutation.

The term “FLT3 pathway inhibitor” or “FLT3 targeting compound” or “FLT3inhibitor” refers to a compound that specifically binds to and inhibitsFLT3, which interferes with the activation of FLT3-mediated signaltransduction pathways and reduces cell proliferation in cancer cellsthat overexpress FLT3. In certain embodiments, a FLT3 inhibitor is acompound that specifically binds to and inhibits wild type FLT3. Incertain embodiments, a FLT3 inhibitor is a compound that specificallybinds to and inhibits mutant FLT3, for example, FLT3-ITD (an internaltandem duplication mutation in the JM domain-coding sequence of the FLT3gene) and/or FLT3-KDM (a missense point mutation at the D835 residue andpoint mutations, deletions and insertions in the codons surrounding D835within a TK domain of FLT3). Exemplary FLT3 inhibitors for use hereininclude, but are not limited to quizartinib (AC220), sunitinib(SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib(CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib(AP24534), ASP2215, KW-2449, famitinib and DCC-2036.

The term “elevated levels of 2HG” means 10%, 20% 30%, 50%, 75%, 100%,200%, 500% or more 2HG is present in a subject that carries a mutantIDH1 allele than is present in a subject that does not carry a mutantIDH1 allele. The term “elevated levels of 2HG” may refer to the amountof 2HG within a cell, within a tumor, within an organ comprising atumor, or within a bodily fluid.

The term “bodily fluid” includes one or more of amniotic fluidsurrounding a fetus, aqueous humour, blood (e.g., blood plasma), serum,Cerebrospinal fluid, cerumen, chyme, Cowper's fluid, female ejaculate,interstitial fluid, lymph, breast milk, mucus (e.g., nasal drainage orphlegm), pleural fluid, pus, saliva, sebum, semen, serum, sweat, tears,urine, vaginal secretion, or vomit.

The terms “inhibit” or “prevent” include both complete and partialinhibition and prevention. An inhibitor may completely or partiallyinhibit the intended target.

The term “subject” is intended to include human and non-human animals.Exemplary human subjects include a human patient (referred to as apatient) having a disorder, e.g., a disorder described herein or anormal subject. The term “non-human animals” of one aspect of theinvention includes all vertebrates, e.g., non-mammals (such as chickens,amphibians, reptiles) and mammals, such as non-human primates,domesticated and/or agriculturally useful animals, e.g., sheep, dog,cat, cow, pig, etc.

The term “treat” means decrease, suppress, attenuate, diminish, arrest,or stabilize the development or progression of a disease/disorder (e.g.,an hematologic malignancy, including an advanced hematologic malignancy,such as acute myelogenous leukemia (AML), myelodysplastic syndrome(MDS), myeloproliferative neoplasms (MPN), chronic myelomonocyticleukemia (CMML), B-acute lymphoblastic leukemias (B-ALL), or lymphoma(e.g., T-cell lymphoma), or a solid tumor, including glioma, melanoma,chondrosarcoma, cholangiocarcinoma (including intrahepaticcholangiocarcinoma (IHCC), prostate cancer, colon cancer, or non-smallcell lung cancer (NSCLC), each characterized by the presence of a mutantallele of IDH1; or acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), chronic myelomonocytic leukemia (CMML), myeloid sarcoma,multiple myeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, or solid tumors, such as glioma, melanoma,chondrosarcoma, or cholangiocarcinoma (e.g., glioma), orangioimmunoblastic T-cell lymphoma (AITL), each characterized by thepresence of a mutant allele of IDH2), lessen the severity of thedisease/disorder or improve the symptoms associated with thedisease/disorder. In one embodiment, the advanced hematologic malignancyis relapsed or refractory. In one embodiment, the solid tumor isrelapsed or refractory.

An amount of a compound, including a pharmaceutically acceptable salt,solvate, tautomer, stereoisomer, isotopologue, prodrug or a polymorphthereof, effective to treat a disorder, or a “therapeutically effectiveamount” or “therapeutically effective dose” refers to an amount of thecompound, including a pharmaceutically acceptable salt, solvate,tautomer, stereoisomer, isotopologue, prodrug, or a polymorph thereof,which is effective, upon single or multiple dose administration to asubject, in treating a cell, or in curing, alleviating, relieving orimproving a subject with a disorder beyond that expected in the absenceof such treatment.

The term “co-administering” as used herein with respect to additionalcancer therapeutic agents means that the additional cancer therapeuticagent may be administered together with a compound provided herein aspart of a single dosage form (such as a composition comprising acompound and a second therapeutic agent as described above) or asseparate, multiple dosage forms. Alternatively, the additional cancertherapeutic agent may be administered prior to, consecutively with, orfollowing the administration of a compound provided herein. In suchcombination therapy treatment, both the compounds provided herein andthe second therapeutic agent(s) are administered by conventionalmethods. The administration of a composition comprising both a compoundprovided herein and a second therapeutic agent, to a subject does notpreclude the separate administration of that same therapeutic agent, anyother second therapeutic agent or any compound provided herein to saidsubject at another time during a course of treatment. The term“co-administering” as used herein with respect to an additional cancertreatment means that the additional cancer treatment may occur prior to,consecutively with, concurrently with or following the administration ofa compound provided herein.

The term “substantially free of other stereoisomers” as used hereinmeans a preparation enriched in a compound having a selectedstereochemistry at one or more selected stereocenters by at least about60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.

The term “enriched” means that at least the designated percentage of apreparation is the compound having a selected stereochemistry at one ormore selected stereocenters.

The term “crystalline” refers to a solid having a highly regularchemical structure. In particular, a crystalline COMPOUND 1 may beproduced as one or more single crystalline forms of COMPOUND 1 and acrystalline COMPOUND 2 may be produced as one or more single crystallineforms of COMPOUND 2. For the purposes of this application, the terms“crystalline form”, “single crystalline form” and “polymorph” aresynonymous; the terms distinguish between crystals that have differentproperties (e.g., different XRPD patterns and/or different DSC scanresults). The term “polymorph” includes pseudopolymorphs, which aretypically different solvates of a material, and thus their propertiesdiffer from one another. Thus, each distinct polymorph andpseudopolymorph of COMPOUND 1 is considered to be a distinct singlecrystalline form herein, and each distinct polymorph and pseudopolymorphof COMPOUND 2 is considered to be a distinct single crystalline formherein.

The term “substantially crystalline” refers to forms that may be atleast a particular weight percent crystalline. Particular weightpercentages are 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%,99.9%, or any percentage between 10% and 100%. In some embodiments,substantially crystalline COMPOUND 1 refers to a COMPOUND 1 that is atleast 70% crystalline. In other embodiments, substantially crystallineCOMPOUND 1 refers to a COMPOUND 1 that is at least 90% crystalline. Insome embodiments, substantially crystalline COMPOUND 2 refers to aCOMPOUND 2 that is at least 70% crystalline. In other embodiments,substantially crystalline COMPOUND 2 refers to a COMPOUND 2 that is atleast 90% crystalline.

The term “isolated” refers to forms that may be at least a particularweight percent of a particular crystalline form of compound. Particularweight percentages are 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%,99.5%, 99.9%, or any percentage between 90% and 100%.

The term “solvate or solvated” means a physical association of acompound, including a crystalline form thereof, of this invention withone or more solvent molecules. This physical association includeshydrogen bonding. In certain instances the solvate will be capable ofisolation, for example when one or more solvent molecules areincorporated in the crystal lattice of the crystalline solid. “Solvateor solvated” encompasses both solution-phase and isolable solvates.Representative solvates include, for example, a hydrate, ethanolates ora methanolate.

The term “hydrate” is a solvate wherein the solvent molecule is H₂O thatis present in a defined stoichiometric amount, and may, for example,include hemihydrate, monohydrate, dihydrate, or trihydrate.

The term “mixture” is used to refer to the combined elements of themixture regardless of the phase-state of the combination (e.g., liquidor liquid/crystalline).

The term “seeding” is used to refer to the addition of a crystallinematerial to initiate recrystallization or crystallization.

The term “antisolvent” is used to refer to a solvent in which compounds,including crystalline forms thereof, are poorly soluble.

The term “pharmaceutically acceptable carrier or adjuvant” refers to acarrier or adjuvant that may be administered to a subject, together witha compound of one aspect of this invention, and which does not destroythe pharmacological activity thereof and is nontoxic when administeredin doses sufficient to deliver a therapeutic amount of the compound.

The term “a pharmaceutically-acceptable salt” as used herein refers tonon-toxic acid or base addition salts of the compound to which the termrefers. Examples of pharmaceutically acceptable salts are discussed inBerge et al., 1977, “Pharmaceutically Acceptable Salts.” J. Pharm. Sci.Vol. 66, pp. 1-19.

The term “about” means approximately, in the region of, roughly, oraround. When the term “about” is used in conjunction with a numericalrange, it modifies that range by extending the boundaries above andbelow the numerical values set forth. In general, the term “about” isused herein to modify a numerical value above and below the stated valueby a variance of 10%.

Compounds

A. Compound 1

In one embodiment, COMPOUND 1 is2-methyl-1-[(4-[6-(trifluoromethyl)pyridin-2-yl]-6-{[2-(trifluoromethyl)pyridin-4-yl]amino}-1,3,5-triazin-2-yl)amino]propan-2-ol,or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof, having thefollowing formula:

COMPOUND 1 may also comprise one or more isotopic substitutions(“Isotopologues”). For example, H may be in any isotopic form, including¹H, ²H (D or deuterium), and ³H (T or tritium); C may be in any isotopicform, including ¹²C, ¹³C, and ¹⁴C; O may be in any isotopic form,including ¹⁶O and ¹⁸O; and the like. For example, COMPOUND 1 is enrichedin a specific isotopic form of H, C and/or O by at least about 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.

COMPOUND 1 in certain embodiments may also be represented in multipletautomeric forms, in such instances, one aspect of the inventionexpressly includes all tautomeric forms of COMPOUND 1 described herein,even though only a single tautomeric form may be represented (e.g.,keto-enol tautomers). All such isomeric forms of COMPOUND 1 areexpressly included herein. Synthesis of COMPOUND 1 is described in USpublished application US-2013-0190287-A1 published Jul. 25, 2013, whichis incorporated by reference in its entirety.

It may be convenient or desirable to prepare, purify, and/or handle acorresponding salt of COMPOUND 1, for example, apharmaceutically-acceptable salt. Examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts.” J. Pharm. Sci. Vol. 66, pp. 1-19.

For example, if COMPOUND 1 is anionic, or has a functional group whichmay be anionic (e.g., —NH— may be —N—⁻), then a salt may be formed witha suitable cation. Examples of suitable inorganic cations include, butare not limited to, alkali metal ions such as Na⁺ and K⁺, alkaline earthcations such as Ca²⁺ and Mg²⁺, and other cations such as Al³⁺. Examplesof some suitable substituted ammonium ions are those derived from:ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

If COMPOUND 1 is cationic, or has a functional group that may becationic (e.g., —NHR may be —NH₂R⁺), then a salt may be formed with asuitable anion. Examples of suitable inorganic anions include, but arenot limited to, those derived from the following inorganic acids:hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,nitrous, phosphoric, and phosphorous.

Examples of suitable organic anions include, but are not limited to,those derived from the following organic acids: 2-acetyoxybenzoic,acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric,edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucoheptonic,gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic,methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic,phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic,succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. In oneembodiment, COMPOUND 1 comprises the mesylate salt of2-methyl-1-[(4-[6-(trifluoromethyl)pyridin-2-yl]-6-{[2-(trifluoromethyl)pyridin-4-yl]amino}-1,3,5-triazin-2-yl)amino]propan-2-ol.Examples of suitable polymeric organic anions include, but are notlimited to, those derived from the following polymeric acids: tannicacid, carboxymethyl cellulose.

COMPOUND 1 for use in the methods and pharmaceutical compositionsprovided herein therefore includes the COMPOUND 1 itself, as well as itspharmaceutically acceptable salts, solvates, tautomers, stereoisomers,isotopologues, prodrugs, metabolites, or polymorphs. Metabolites ofCOMPOUND 1 are disclosed in patent application publicationWO2015/006592, which is incorporated herein by reference in itsentirety. COMPOUND 1 provided herein may be modified and converted to aprodrug by appending appropriate functionalities to enhance selectedbiological properties, e.g., targeting to a particular tissue. Suchmodifications (i.e., prodrugs) are known in the art and include thosewhich increase biological penetration into a given biologicalcompartment (e.g., blood, lymphatic system, central nervous system),increase oral availability, increase solubility to allow administrationby injection, alter metabolism and alter rate of excretion. Examples ofprodrugs include esters (e.g., phosphates, amino acid (e.g., valine)esters), carbamates and other pharmaceutically acceptable derivatives,which, upon administration to a subject, are capable of providing activecompounds.

It has been found that COMPOUND 1 can exist in a variety of solid forms.In one embodiment, provided herein are solid forms that include neatcrystal forms. In another embodiment, provided herein are solid formsthat include solvated forms and amorphous forms. The present disclosureprovides certain solid forms of COMPOUND 1. In certain embodiments, thepresent disclosure provides compositions comprising COMPOUND 1 in a formdescribed herein. In some embodiments of provided compositions, COMPOUND1 is present as a mixture of one or more solid forms; in someembodiments of provided compositions, COMPOUND 1 is present in a singleform.

In one embodiment, COMPOUND 1 is a single crystalline form, or any oneof the single crystalline forms described herein. Synthesis ofcrystalline forms of COMPOUND 1 is described in the internationalapplication publication WO 2015/017821 published Feb. 5, 2015 and theU.S. provisional application Ser. No. 61/112,127, filed Feb. 4, 2015,both incorporated by reference herein in their entireties. Also providedare pharmaceutical compositions comprising at least one pharmaceuticallyacceptable carrier or diluent; and COMPOUND 1, wherein COMPOUND 1 is asingle crystalline form, or any one of the crystalline forms beingdescribed herein. Also provided are uses of COMPOUND 1, wherein COMPOUND1 is a single crystalline form, or any one of the single crystallineforms described herein, to prepare a pharmaceutical composition.

Provided herein is an assortment of characterizing information todescribe the crystalline forms of COMPOUND 1. It should be understood,however, that not all such information is required for one skilled inthe art to determine that such particular form is present in a givencomposition, but that the determination of a particular form can beachieved using any portion of the characterizing information that oneskilled in the art would recognize as sufficient for establishing thepresence of a particular form, e.g., even a single distinguishing peakcan be sufficient for one skilled in the art to appreciate that suchparticular form is present.

In one embodiment, at least a particular percentage by weight ofCOMPOUND 1 is crystalline. Particular weight percentages may be 10%,20%, 30%, 40%, 50%, 60%, 70%, 75%,

80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99%, 99.5%, 99.9%, or any percentage between 10% and 100%. When aparticular percentage by weight of COMPOUND 1 is crystalline, theremainder of COMPOUND 1 is the amorphous form of COMPOUND 1.Non-limiting examples of crystalline COMPOUND 1 include a singlecrystalline form of compound 1 or a mixture of different singlecrystalline forms. In some embodiments, COMPOUND 1 is at least 90% byweight crystalline. In some other embodiments, COMPOUND 1 is at least95% by weight crystalline.

In another embodiment, a particular percentage by weight of thecrystalline COMPOUND 1 is a specific single crystalline form or acombination of single crystalline forms. Particular weight percentagesmay be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or anypercentage between 10% and 100%. In another embodiment, COMPOUND 1 is atleast 90% by weight of a single crystalline form. In another embodiment,COMPOUND 1 is at least 95% by weight of a single crystalline form.

In the following description of COMPOUND 1, embodiments of the inventionmay be described with reference to a particular crystalline form ofCOMPOUND 1, as characterized by one or more properties as discussedherein. The descriptions characterizing the crystalline forms may alsobe used to describe the mixture of different crystalline forms that maybe present in a crystalline COMPOUND 1. However, the particularcrystalline forms of COMPOUND 1 may also be characterized by one or moreof the characteristics of the crystalline form as described herein, withor without regard to referencing a particular crystalline form.

The crystalline forms are further illustrated by the detaileddescriptions and illustrative examples given below. The XRPD peaksdescribed in Tables 1 to 6 may vary by ±0.2° depending upon theinstrument used to obtain the data. The intensity of the XRPD peaksdescribed in Tables 1 to 6 may vary by 10%.

Form 1

In one embodiment, a single crystalline form, Form 1, of COMPOUND 1 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 1, and data shown in Table 1 obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 1, as shown in Table 1. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine of the peaks shown in Table 1.

TABLE 1 Angle 2-Theta° Intensity % 6.7 42.2 8.9 61.8 9.1 41.9 13.0 46.716.4 33.2 18.9 100.0 21.4 27.3 23.8 49.2 28.1 47.5

In another embodiment, Form 1 can be characterized by the peaksidentified at 2θ angles of 8.9, 13.0, 18.9, 23.8, and 28.1°. In anotherembodiment, Form 1 can be characterized by the peaks identified at 2θangles of 8.9, 18.9, and 23.8°.

Form 2

In one embodiment, a single crystalline form, Form 2, of COMPOUND 1 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 2, and data shown in Table 2, obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 2, as shown in Table 2. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine of the peaks shown in Table 2.

TABLE 2 Angle 2-Theta° Intensity % 8.4 65.2 12.7 75.5 16.9 57.9 17.169.4 17.7 48.6 19.2 100.0 23.0 69.7 23.3 61.1 24.2 87.3

In another embodiment, Form 2 can be characterized by the peaksidentified at 2θ angles of 12.7, 17.1, 19.2, 23.0, and 24.2°. In anotherembodiment, Form 2 can be characterized by the peaks identified at 2θangles of 12.7, 19.2, and 24.2°.

Form 3

In one embodiment, a single crystalline form, Form 3, of COMPOUND 1 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 3, and data shown in Table 3, obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 3, as shown in Table 3. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine of the peaks shown in Table 3.

TABLE 3 Angle 2-Theta° Intensity % 6.8 35.5 10.1 30.7 10.6 53.1 13.646.0 14.2 63.8 17.2 26.4 18.4 34.0 19.2 100.0 23.5 3.8

In another embodiment, Form 3 can be characterized by the peaksidentified at 2θ angles of 6.8, 10.6, 13.6, 14.2, and 19.2°. In anotherembodiment, Form 3 can be characterized by the peaks identified at 2θangles of 10.6, 14.2, and 19.2°.

Form 4

In one embodiment, a single crystalline form, Form 4, of COMPOUND 1 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 4, and data shown in Table 4, obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 4, as shown in Table 4. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine of the peaks shown in Table 4.

TABLE 4 Angle 2-Theta° Intensity % 7.2 53.3 10.1 26.7 11.5 20.5 13.6100.0 18.5 72.0 19.3 46.9 20.3 39.4 21.9 55.4 23.5 77.5

In another embodiment, Form 4 can be characterized by the peaksidentified at 2θ angles of 7.2, 13.6, 18.5, 19.3, 21.9, and 23.5°. Inanother embodiment, Form 4 can be characterized by the peaks identifiedat 2θ angles of 13.6, 18.5, and 23.5°.

Form 5

In one embodiment, a single crystalline form, Form 5, of COMPOUND 1 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 5, and data shown in Table 5, obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 5, as shown in Table 5. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine of the peaks shown in Table 5.

TABLE 5 Angle 2-Theta° Intensity % 6.4 45.4 8.4 84.0 9.8 100.0 16.1 26.016.9 22.7 17.8 43.6 19.7 40.4 21.1 20.5 26.1 15.9

In another embodiment, Form 5 can be characterized by the peaksidentified at 2θ angles of 6.4, 8.4, 9.8, 17.8, and 19.7°. In anotherembodiment, Form 5 can be characterized by the peaks identified at 2θangles of 8.4 and 9.8°.

Form 6

In one embodiment, a single crystalline form, Form 6, of COMPOUND 1 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 15, and data shown in Table 6, obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 6, as shown in Table 6. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight of the peaks shown in Table 6.

TABLE 6 Angle 2-Theta° Intensity % 8.1 97.9 11.4 24.9 14.1 51.5 15.228.4 16.4 85.0 17.3 100.0 20.5 54.7 24.1 88.7

In another embodiment, Form 6 can be characterized by the peaksidentified at 2θ angles of 8.1, 14.1, 16.4, 17.3, 20.5, and 24.1°. Inanother embodiment, Form 6 can be characterized by the peaks identifiedat 2θ angles of 8.1, 16.4, 17.3, and 24.1°.

B. Compound 2

COMPOUND 2 is(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide,a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, or a polymorph thereof. COMPOUND 2 has thefollowing chemical structure:

COMPOUND 2 may also comprise one or more isotopic substitutions. Forexample, H may be in any isotopic form (“Isotopologues”), including ¹H,²H (D or deuterium), and ³H (T or tritium); C may be in any isotopicform, including ¹²C, ¹³C, and ¹⁴C; O may be in any isotopic form,including ¹⁶O and ¹⁸O; and the like. For example, COMPOUND 2 is enrichedin a specific isotopic form of H, C and/or O by at least about 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%.

COMPOUND 2 in certain embodiments may also be represented in multipletautomeric forms, in such instances, one aspect of the inventionexpressly includes all tautomeric forms of COMPOUND 2 described herein,even though only a single tautomeric form may be represented (e.g.,keto-enol tautomers). All such isomeric forms of COMPOUND 2 areexpressly included herein. Synthesis of COMPOUND 2 is described in USpublished application US-2013-0190249-A1 published Jul. 25, 2013, whichis incorporated by reference in its entirety.

It may be convenient or desirable to prepare, purify, and/or handle acorresponding salt of COMPOUND 2, for example, apharmaceutically-acceptable salt. Examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts.” J. Pharm. Sci. Vol. 66, pp. 1-19.

For example, if COMPOUND 2 is anionic, or has a functional group whichmay be anionic (e.g., —NH— may be —N—⁻), then a salt may be formed witha suitable cation. Examples of suitable inorganic cations include, butare not limited to, alkali metal ions such as Na⁺ and K⁺, alkaline earthcations such as Ca²⁺ and Mg²⁺, and other cations such as Al³⁺. Examplesof some suitable substituted ammonium ions are those derived from:ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

If COMPOUND 2 is cationic, or has a functional group that may becationic (e.g., —NHR may be —NH₂R⁺), then a salt may be formed with asuitable anion. Examples of suitable inorganic anions include, but arenot limited to, those derived from the following inorganic acids:hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,nitrous, phosphoric, and phosphorous.

Examples of suitable organic anions include, but are not limited to,those derived from the following organic acids: 2-acetyoxybenzoic,acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric,edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucoheptonic,gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic,methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic,phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic,succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examplesof suitable polymeric organic anions include, but are not limited to,those derived from the following polymeric acids: tannic acid,carboxymethyl cellulose.

COMPOUND 2 for use in the methods and pharmaceutical compositionsprovided herein therefore includes COMPOUND 2 itself, as well as itspharmaceutically acceptable salts, solvates, tautomers, stereoisomers,isotopologues, prodrugs or polymorphs. COMPOUND 2 provided herein may bemodified and converted to a prodrug by appending appropriatefunctionalities to enhance selected biological properties, e.g.,targeting to a particular tissue. Such modifications (i.e., prodrugs)are known in the art and include those which increase biologicalpenetration into a given biological compartment (e.g., blood, lymphaticsystem, central nervous system), increase oral availability, increasesolubility to allow administration by injection, alter metabolism andalter rate of excretion. Examples of prodrugs include esters (e.g.,phosphates, amino acid (e.g., valine) esters), carbamates and otherpharmaceutically acceptable derivatives, which, upon administration to asubject, are capable of providing active compounds.

It has been found that COMPOUND 2 can exist in a variety of solid forms.In one embodiment, provided herein are solid forms that include neatcrystal forms. In another embodiment, provided herein are solid formsthat include solvated forms and amorphous forms. The present disclosureprovides certain solid forms of COMPOUND 2. In certain embodiments, thepresent disclosure provides compositions comprising COMPOUND 2 in a formdescribed herein. In some embodiments of provided compositions, COMPOUND2 is present as a mixture of one or more solid forms; in someembodiments of provided compositions, COMPOUND 2 is present in a singleform.

In one embodiment, COMPOUND 2 is a single crystalline form, or any oneof the single crystalline forms described herein. Synthesis ofcrystalline forms of COMPOUND 2 is described in internationalapplication publications WO 2015/138837 and WO 2015/138839, bothpublished Sep. 17, 2015, both incorporated by reference herein in theirentireties. Also provided are pharmaceutical compositions comprising atleast one pharmaceutically acceptable carrier or diluent; and COMPOUND2, wherein COMPOUND 2 is a single crystalline form, or any one of thecrystalline forms being described herein. Also provided are uses ofCOMPOUND 2, wherein COMPOUND 2 is a single crystalline form, or any oneof the single crystalline forms described herein, to prepare apharmaceutical composition.

Provided herein is an assortment of characterizing information todescribe the crystalline forms of COMPOUND 2. It should be understood,however, that not all such information is required for one skilled inthe art to determine that such particular form is present in a givencomposition, but that the determination of a particular form can beachieved using any portion of the characterizing information that oneskilled in the art would recognize as sufficient for establishing thepresence of a particular form, e.g., even a single distinguishing peakcan be sufficient for one skilled in the art to appreciate that suchparticular form is present.

In one embodiment, at least a particular percentage by weight ofCOMPOUND 2 is crystalline. Particular weight percentages may be 10%,20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or any percentagebetween 10% and 100%. When a particular percentage by weight of COMPOUND2 is crystalline, the remainder of COMPOUND 2 is the amorphous form ofCOMPOUND 2. Non-limiting examples of crystalline COMPOUND 2 include asingle crystalline form of compound 1 or a mixture of different singlecrystalline forms. In some embodiments, COMPOUND 2 is at least 90% byweight crystalline. In some other embodiments, COMPOUND 2 is at least95% by weight crystalline.

In another embodiment, a particular percentage by weight of thecrystalline COMPOUND 2 is a specific single crystalline form or acombination of single crystalline forms. Particular weight percentagesmay be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or anypercentage between 10% and 100%. In another embodiment, COMPOUND 2 is atleast 90% by weight of a single crystalline form. In another embodiment,COMPOUND 2 is at least 95% by weight of a single crystalline form.

In the following description of COMPOUND 2, embodiments of the inventionmay be described with reference to a particular crystalline form ofCOMPOUND 2, as characterized by one or more properties as discussedherein. The descriptions characterizing the crystalline forms may alsobe used to describe the mixture of different crystalline forms that maybe present in a crystalline COMPOUND 2. However, the particularcrystalline forms of COMPOUND 2 may also be characterized by one or moreof the characteristics of the crystalline form as described herein, withor without regard to referencing a particular crystalline form.

The crystalline forms are further illustrated by the detaileddescriptions and illustrative examples given below. The XRPD peaksdescribed in Tables 1 to 2 may vary by ±0.2° depending upon theinstrument used to obtain the data. The intensity of the XRPD peaksdescribed in Tables 1 to 2 may vary by 10%.

Form I

In one embodiment, a single crystalline form, Form I, of COMPOUND 2 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 7, and data shown in Table 7, obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 7, as shown in Table 7. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine of the peaks shown in Table 7.

TABLE 7 Angle 2-Theta° Intensity % 8.6 90.3 13.2 60.0 15.6 85.5 18.572.5 19.6 31.5 20.6 71.6 21.6 100.0 26.4 64.2 27.3 45.6

In another embodiment, Form I can be characterized by the peaksidentified at 2θ angles of 8.6, 15.6, 18.5, 20.6, 21.6, and 26.4°. Inanother embodiment, Form I can be characterized by the peaks identifiedat 2θ angles of 8.6, 15.6, 18.5, and 21.6°.

In another embodiment, Form I can be characterized by the differentialscanning calorimetry profile (DSC) shown in FIG. 8. The DSC graph plotsthe heat flow as a function of temperature from a sample, thetemperature rate change being about 10° C./min. The profile ischaracterized by an endothermic transition with an onset temperature ofabout 140.1° C. with a melt at about 149.9° C.

In another embodiment, Form I can be characterized by thermalgravimetric analysis (TGA) shown in FIG. 9. The TGA profile graphs thepercent loss of weight of the sample as a function of temperature, thetemperature rate change being about 10° C./min. The weight lossrepresents a loss of about 0.44% of the weight of the sample as thetemperature is changed from about 29.0° C. to 125.0° C.

Form II

In one embodiment, a single crystalline form, Form II, of the COMPOUND 2is characterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 10, and data shown in Table 8, obtained using CuKα radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 10, as shown in Table 8. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine or ten of the peaks shown in Table 8.

TABLE 8 Angle 2-Theta° Intensity % 9.8 85.6 11.6 100.0 14.9 11.4 16.515.3 19.6 75.2 20.1 7.3 22.5 32.6 23.0 69.4 25.0 8.9 31.4 22.0

In another embodiment, Form II can be characterized by the peaksidentified at 2θ angles of 9.8, 11.6, 19.6, 22.5, 23.0, and 31.4°. Inanother embodiment, Form II can be characterized by the peaks identifiedat 2θ angles of 9.8, 11.6, 19.6, and 23.0°.

In another embodiment, Form II can be characterized by the differentialscanning calorimetry profile (DSC) shown in FIG. 11. The DSC graph plotsthe heat flow as a function of temperature from a sample, thetemperature rate change being about 10° C./min. The profile ischaracterized by an endothermic transition with an onset temperature ofabout 62.7° C. with a melt at about 72.5° C., and an endothermictransition with an onset temperature of about 145.6° C. with a melt atabout 153.6° C.

In another embodiment, Form II can be characterized by thermalgravimetric analysis (TGA) shown in FIG. 12. The TGA profile graphs thepercent loss of weight of the sample as a function of temperature, thetemperature rate change being about 10° C./min. The weight lossrepresents a loss of about 0.57% of the weight of the sample as thetemperature is changed from about 29.3° C. to 170.3° C.

Other embodiments are directed to a single crystalline form of COMPOUND2 characterized by a combination of the aforementioned characteristicsof any of the single crystalline forms discussed herein. Thecharacterization may be by any combination of one or more of the XRPD,TGA, and DSC described for a particular polymorph. For example, thesingle crystalline form of COMPOUND 2 may be characterized by anycombination of the XRPD results regarding the position of the majorpeaks in a XRPD scan; and/or any combination of one or more ofparameters derived from data obtained from a XRPD scan. The singlecrystalline form of COMPOUND 2 may also be characterized by TGAdeterminations of the weight loss associated with a sample over adesignated temperature range; and/or the temperature at which aparticular weight loss transition begins. DSC determinations of thetemperature associated with the maximum heat flow during a heat flowtransition and/or the temperature at which a sample begins to undergo aheat flow transition may also characterize the crystalline form. Weightchange in a sample and/or change in sorption/desorption of water permolecule of COMPOUND 2 as determined by water sorption/desorptionmeasurements over a range of relative humidity (e.g., 0% to 90%) mayalso characterize a single crystalline form of COMPOUND 2.

FLT3 Targeting Compounds

In one embodiment, the methods provided herein compriseco-administration of one or more second agent, wherein the second agentis a FLT3 targeting agent.

In one aspect, the second agent specifically binds to and inhibits FLT3,which interferes with the activation of FLT3-mediated signaltransduction pathways and reduces cell proliferation in cancer cellsthat overexpress FLT3. Exemplary second agents that target FLT3 aredescribed by Hitoshi Kiyo Nagoya J Med Sci. 2015 February; 77(1-2): 7-17and Fathi et al., Am J Blood Res 2011; 1(2):175-189, incorporated byreference herein.

In certain embodiments, a FLT3 inhibitor is a compound that specificallybinds to and inhibits wild type FLT3. In certain embodiments, a FLT3inhibitor is a compound that specifically binds to and inhibits mutantFLT3. In certain embodiments, a FLT3 inhibitor is a compound thatspecifically binds to and inhibits FLT3-ITD. In certain embodiments, aFLT3 inhibitor is a compound that specifically binds to and inhibitsFLT3-KDM.

In certain embodiments, the second agent is a FLT3 inhibitor selectedfrom quizartinib (AC220), sunitinib (SU11248), sorafenib (BAY 43-9006),midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596),PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449,famitinib and DCC-2036.

In certain embodiments, the second agent is a FLT3 inhibitor selectedfrom quizartinib (AC220), sunitinib (SU11248), sorafenib (BAY 43-9006)and midostaurin (PKC412).

Compositions and Routes of Administration

In one embodiment, provided herein is a pharmaceutical compositioncomprising a therapeutically effective amount of a mutant IDH2inhibitor. In one embodiment, the mutant IDH1 inhibitor is COMPOUND 1.

In one embodiment, provided herein is a pharmaceutical compositioncomprising a therapeutically effective amount of a mutant IDH1inhibitor. In one embodiment, the mutant IDH1 inhibitor is COMPOUND 2.

In one embodiment, the compounds utilized in the methods provided hereinmay be formulated together with a pharmaceutically acceptable carrier oradjuvant into pharmaceutically acceptable compositions prior to beadministered to a subject. In another embodiment, such pharmaceuticallyacceptable compositions further comprise additional therapeutic agentsin amounts effective for achieving a modulation of disease or diseasesymptoms, including those described herein.

Pharmaceutically acceptable carriers, adjuvants and vehicles that may beused in the pharmaceutical compositions of one aspect of this inventioninclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, self-emulsifying drug delivery systems (SEDDS) suchas d-α-tocopherol polyethyleneglycol 1000 succinate, surfactants used inpharmaceutical dosage forms such as Tweens or other similar polymericdelivery matrices, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat. Cyclodextrins such as α-, β-, and γ-cyclodextrin, orchemically modified derivatives such as hydroxyalkylcyclodextrins,including 2- and 3-hydroxypropyl-β-cyclodextrins, or other solubilizedderivatives may also be advantageously used to enhance delivery ofCOMPOUND 1 or COMPOUND 2 described herein.

In one embodiment, the pharmaceutical composition comprises COMPOUND 1and an excipient. In one embodiment, the pharmaceutical composition thatcomprises COMPOUND 1 and an excipient, is for oral administration. Inone embodiment, the excipient is a diluent, a binder, a disintegrant, awetting agent, a stabilizer, a glidant, or a lubricant.

In one embodiment, the pharmaceutical composition comprises COMPOUND 2and an excipient. In one embodiment, the pharmaceutical composition thatcomprises COMPOUND 2 and an excipient, is for oral administration. Inone embodiment, the excipient is a diluent, a binder, a disintegrant, awetting agent, a stabilizer, a glidant, or a lubricant.

In one embodiment, the diluent is a microcrystalline cellulose.

In one embodiment, the binder is a hydroxypropyl cellulose.

In one embodiment, the disintegrant is sodium starch glycolate.

In one embodiment, the wetting agent is sodium lauryl sulfate.

In one embodiment, the stabilizer is hypromellose acetate succinate.

In one embodiment, the glidant is colloidal silicon dioxide.

In one embodiment, the lubricant is magnesiun stearate.

In one embodiment, the pharmaceutical composition comprises COMPOUND 1or COMPOUND 2 and an excipient. In one embodiment, the pharmaceuticalcomposition that comprises COMPOUND 1 or COMPOUND 2 and an excipient, isfor oral administration.

Oral delivery formats for COMPOUND 1 or COMPOUND 2 include, but are notlimited to, tablets, capsules, caplets, solutions, suspensions, andsyrups, and may also comprise a plurality of granules, beads, powders orpellets that may or may not be encapsulated. Such formats may also bereferred to herein as the “drug core” which contains COMPOUND 1 orCOMPOUND 2.

Particular embodiments herein provide solid oral dosage forms that aretablets or capsules. In certain embodiments, the formulation is a tabletcomprising COMPOUND 1 or COMPOUND 2. In certain embodiments, theformulation is a capsule comprising COMPOUND 1 or COMPOUND 2. In certainembodiments, the tablets or capsules provided herein optionally compriseone or more excipients, such as, for example, glidants, diluents,lubricants, colorants, disintegrants, granulating agents, bindingagents, polymers, and coating agents. In certain embodiments, theformulation is an immediate release tablet. In certain embodiments, theformulation is a controlled release tablet releasing the activepharmaceutical ingredient (API), e.g., substantially in the stomach. Incertain embodiments, the formulation is a hard gelatin capsule. Incertain embodiments, the formulation is a soft gelatin capsule. Incertain embodiments, the capsule is a hydroxypropyl methylcellulose(HPMC) capsule. In certain embodiments, the formulation is an immediaterelease capsule. In certain embodiments, the formulation is an immediateor controlled release capsule releasing the API, e.g., substantially inthe stomach. In certain embodiments, the formulation is a rapidlydisintegrating tablet that dissolves substantially in the mouthfollowing administration. In certain embodiments, embodiments hereinencompass the use of COMPOUND 1 for the preparation of a pharmaceuticalcomposition for treating a malignancy, characterized by the presence ofa mutant allele of IDH2, wherein the composition is prepared for oraladministration. In certain embodiments, embodiments herein encompass theuse of COMPOUND 2 for the preparation of a pharmaceutical compositionfor treating a malignancy, characterized by the presence of a mutantallele of IDH1, wherein the composition is prepared for oraladministration.

Particular embodiments herein provide pharmaceutical formulations (e.g.,immediate release oral formulations and/or formulations that release theAPI substantially in the stomach) comprising COMPOUND 1 or COMPOUND 2that achieve a particular AUC value (e.g., AUC(0-t) or AUC(0-∞)) in thesubject (e.g., human) to which the formulation is orally administered.Particular embodiments provide oral formulations that achieve an AUCvalue of at least about 25 ng-hr/mL, at least about 50 ng-hr/mL, atleast about 75 ng-hr/mL, at least about 100 ng-hr/mL, at least about 150ng-hr/mL, at least about 200 ng-hr/mL, at least about 250 ng-hr/mL, atleast about 300 ng-hr/mL, at least about 350 ng-hr/mL, at least about400 ng-hr/mL, at least about 450 ng-hr/mL, at least about 500 ng-hr/mL,at least about 550 ng-hr/mL, at least about 600 ng-hr/mL, at least about650 ng-hr/mL, at least about 700 ng-hr/mL, at least about 750 ng-hr/mL,at least about 800 ng-hr/mL, at least about 850 ng-hr/mL, at least about900 ng-hr/mL, at least about 950 ng-hr/mL, at least about 1000 ng-hr/mL,at least about 1100 ng-hr/mL, at least about 1200 ng-hr/mL, at leastabout 1300 ng-hr/mL, at least about 1400 ng-hr/mL, at least about 1500ng-hr/mL, at least about 1600 ng-hr/mL, at least about 1700 ng-hr/mL, atleast about 1800 ng-hr/mL, at least about 1900 ng-hr/mL, at least about2000 ng-hr/mL, at least about 2250 ng-hr/mL, or at least about 2500ng-hr/mL. In particular embodiments, the AUC determination is obtainedfrom a time-concentration pharmacokinetic profile obtained from theblood samples of animals or human volunteers following dosing.

Particular embodiments herein provide pharmaceutical formulations (e.g.,immediate release oral formulations and/or formulations that release theAPI substantially in the stomach) comprising COMPOUND 1 or COMPOUND 2that achieve a particular maximum plasma concentration (“Cmax”) in thesubject to which the formulation is orally administered. Particularembodiments provide oral formulations that achieve a Cmax of COMPOUND 1or COMPOUND 2 of at least about 25 ng/mL, at least about 50 ng/mL, atleast about 75 ng/mL, at least about 100 ng/mL, at least about 150ng/mL, at least about 200 ng/mL, at least about 250 ng/mL, at leastabout 300 ng/mL, at least about 350 ng/mL, at least about 400 ng/mL, atleast about 450 ng/mL, at least about 500 ng/mL, at least about 550ng/mL, at least about 600 ng/mL, at least about 650 ng/mL, at leastabout 700 ng/mL, at least about 750 ng/mL, at least about 800 ng/mL, atleast about 850 ng/mL, at least about 900 ng/mL, at least about 950ng/mL, at least about 1000 ng/mL, at least about 1100 ng/mL, at leastabout 1200 ng/mL, at least about 1300 ng/mL, at least about 1400 ng/mL,at least about 1500 ng/mL, at least about 1600 ng/mL, at least about1700 ng/mL, at least about 1800 ng/mL, at least about 1900 ng/mL, atleast about 2000 ng/mL, at least about 2250 ng/mL, or at least about2500 ng/mL.

Particular embodiments herein provide pharmaceutical formulations (e.g.,immediate release oral formulations and/or formulations that release theAPI substantially in the stomach) comprising COMPOUND 1 or COMPOUND 2that achieve a particular time to maximum plasma concentration (“Tmax”)in the subject to which the formulation is orally administered.Particular embodiments provide oral formulations that achieve a Tmax ofthe cytidine analog of less than about 10 min., less than about 15 min.,less than about 20 min., less than about 25 min., less than about 30min., less than about 35 min., less than about 40 min., less than about45 min., less than about 50 min., less than about 55 min., less thanabout 60 min., less than about 65 min., less than about 70 min., lessthan about 75 min., less than about 80 min., less than about 85 min.,less than about 90 min., less than about 95 min., less than about 100min., less than about 105 min., less than about 110 min., less thanabout 115 min., less than about 120 min., less than about 130 min., lessthan about 140 min., less than about 150 min., less than about 160 min.,less than about 170 min., less than about 180 min., less than about 190min., less than about 200 min., less than about 210 min., less thanabout 220 min., less than about 230 min., or less than about 240 min. Inparticular embodiments, the Tmax value is measured from the time atwhich the formulation is orally administered.

Particular embodiments herein provide oral dosage forms comprisingCOMPOUND 1 or COMPOUND 2 wherein the oral dosage forms have an entericcoating. Particular embodiments provide a permeable or partly permeable(e.g., “leaky”) enteric coating with pores. In particular embodiments,the permeable or partly permeable enteric-coated tablet releasesCOMPOUND 1 or COMPOUND 2 in an immediate release manner substantially inthe stomach.

Provided herein are dosage forms designed to maximize the absorptionand/or efficacious delivery of COMPOUND 1 or COMPOUND 2, upon oraladministration, e.g., for release substantially in the stomach.Accordingly, certain embodiments herein provide a solid oral dosage formof COMPOUND 1 or COMPOUND 2 using pharmaceutical excipients designed forimmediate release of the API upon oral administration, e.g.,substantially in the stomach. Particular immediate release formulationscomprise a specific amount of COMPOUND 1 or COMPOUND 2 and optionallyone or more excipients. In certain embodiments, the formulation may bean immediate release tablet or an immediate release capsule (such as,e.g., an HPMC capsule).

Provided herein are methods of making the formulations provided hereincomprising COMPOUND 1 or COMPOUND 2 provided herein (e.g., immediaterelease oral formulations and/or formulations that release the APIsubstantially in the stomach). In particular embodiments, theformulations provided herein may be prepared using conventional methodsknown to those skilled in the field of pharmaceutical formulation, asdescribed, e.g., in pertinent textbooks. See, e.g., REMINGTON, THESCIENCE AND PRACTICE OF PHARMACY, 20th Edition, Lippincott Williams &Wilkins, (2000); ANSEL et al., PHARMACEUTICAL DOSAGE FORMS AND DRUGDELIVERY SYSTEMS, 7th Edition, Lippincott Williams & Wilkins, (1999);GIBSON, PHARMACEUTICAL PREFORMULATION AND FORMULATION, CRC Press (2001).

In particular embodiments, formulations provided herein (e.g., immediaterelease oral formulations, formulations that release the APIsubstantially in the stomach, or rapidly disintegrating formulationsthat dissolve substantially in the mouth) comprise COMPOUND 1 orCOMPOUND 2 in a specific amount. In particular embodiments, the specificamount of COMPOUND 1 or COMPOUND 2 in the formulation is, e.g., about 10mg. In one embodiment, the specific amount is about 20 mg. In oneembodiment, the specific amount is about 40 mg. In one embodiment, thespecific amount is about 60 mg. In one embodiment, the specific amountis about 80 mg. In one embodiment, the specific amount is about 100 mg.In one embodiment, the specific amount is about 120 mg. In oneembodiment, the specific amount is about 140 mg. In one embodiment, thespecific amount is about 160 mg. In one embodiment, the specific amountis about 180 mg. In one embodiment, the specific amount is about 200 mg.In one embodiment, the specific amount is about 220 mg. In oneembodiment, the specific amount is about 240 mg. In one embodiment, thespecific amount is about 260 mg. In one embodiment, the specific amountis about 280 mg. In one embodiment, the specific amount is about 300 mg.In one embodiment, the specific amount is about 320 mg. In oneembodiment, the specific amount is about 340 mg. In one embodiment, thespecific amount is about 360 mg. In one embodiment, the specific amountis about 380 mg. In one embodiment, the specific amount is about 400 mg.In one embodiment, the specific amount is about 420 mg. In oneembodiment, the specific amount is about 440 mg. In one embodiment, thespecific amount is about 460 mg. In one embodiment, the specific amountis about 480 mg. In one embodiment, the specific amount is about 500 mg.In one embodiment, the specific amount is about 600 mg. In oneembodiment, the specific amount is about 700 mg. In one embodiment, thespecific amount is about 800 mg. In one embodiment, the specific amountis about 900 mg. In one embodiment, the specific amount is about 1000mg. In one embodiment, the specific amount is about 1100 mg. In oneembodiment, the specific amount is about 1200 mg. In one embodiment, thespecific amount is about 1300 mg. In one embodiment, the specific amountis about 1400 mg. In one embodiment, the specific amount is about 1500mg. In one embodiment, the specific amount is about 1600 mg. In oneembodiment, the specific amount is about 1700 mg. In one embodiment, thespecific amount is about 1800 mg. In one embodiment, the specific amountis about 1900 mg. In one embodiment, the specific amount is about 2000mg. In one embodiment, the specific amount is about 2100 mg. In oneembodiment, the specific amount is about 2200 mg. In one embodiment, thespecific amount is about 2300 mg. In one embodiment, the specific amountis about 2400 mg. In one embodiment, the specific amount is about 2500mg. In one embodiment, the specific amount is about 3000 mg. In oneembodiment, the specific amount is about 4000 mg. In one embodiment, thespecific amount is about 5000 mg.

In certain embodiments, the formulation is a tablet, wherein the tabletis manufactured using standard, art-recognized tablet processingprocedures and equipment. In certain embodiments, the method for formingthe tablets is direct compression of a powdered, crystalline and/orgranular composition comprising COMPOUND 1 or COMPOUND 2 alone or incombination with one or more excipients, such as, for example, carriers,additives, polymers, or the like. In certain embodiments, as analternative to direct compression, the tablets may be prepared using wetgranulation or dry granulation processes. In certain embodiments, thetablets are molded rather than compressed, starting with a moist orotherwise tractable material. In certain embodiments, compression andgranulation techniques are used.

In certain embodiments, the formulation is a capsule, wherein thecapsules may be manufactured using standard, art-recognized capsuleprocessing procedures and equipments. In certain embodiments, softgelatin capsules may be prepared in which the capsules contain a mixtureof COMPOUND 1 or COMPOUND 2 and vegetable oil or non-aqueous, watermiscible materials such as, for example, polyethylene glycol and thelike. In certain embodiments, hard gelatin capsules may be preparedcontaining granules of COMPOUND 1 or COMPOUND 2 in combination with asolid pulverulent carrier, such as, for example, lactose, saccharose,sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulosederivatives, or gelatin. In certain embodiments, a hard gelatin capsuleshell may be prepared from a capsule composition comprising gelatin anda small amount of plasticizer such as glycerol. In certain embodiments,as an alternative to gelatin, the capsule shell may be made of acarbohydrate material. In certain embodiments, the capsule compositionmay additionally include polymers, colorings, flavorings and opacifiersas required. In certain embodiments, the capsule comprises HPMC.

In certain embodiments, the formulation of COMPOUND 1 or COMPOUND 2 isprepared using aqueous solvents without causing significant hydrolyticdegradation of the compounds. In particular embodiments, the formulationof COMPOUND 1 or COMPOUND 2 is a tablet which contains a coating appliedto the drug core using aqueous solvents without causing significanthydrolytic degradation of the compound in the formulation. In certainembodiments, water is employed as the solvent for coating the drug core.In certain embodiments, the oral dosage form of COMPOUND 1 or COMPOUND 2is a tablet containing a film coat applied to the drug core usingaqueous solvents. In particular embodiments, water is employed as thesolvent for film-coating. In particular embodiments, the tabletcontaining COMPOUND 1 or COMPOUND 2 is film-coated using aqueoussolvents without effecting degradation of the pharmaceuticalcomposition. In particular embodiments, water is used as the filmcoating solvent without effecting degradation of the pharmaceuticalcomposition. In certain embodiments, an oral dosage form comprisingCOMPOUND 1 or COMPOUND 2 and an aqueous film coating effects immediatedrug release upon oral delivery. In certain embodiments, the oral dosageform comprising COMPOUND 1 or COMPOUND 2 and an aqueous film coatingeffects controlled drug release to the upper gastrointestinal tract,e.g., the stomach, upon oral administration. In particular embodiments,a tablet with an aqueous-based film coating comprises COMPOUND 1 orCOMPOUND 2 as the API.

In certain embodiments, provided herein is a controlled releasepharmaceutical formulation for oral administration of COMPOUND 1 orCOMPOUND 2 substantially in the stomach, comprising: a) a specificamount of COMPOUND 1 or COMPOUND 2; b) a drug release controllingcomponent for controlling the release of COMPOUND 1 or COMPOUND 2substantially in the upper gastrointestinal tract, e.g., the stomach;and c) optionally one or more excipients. In certain embodiments, theoral dosage form comprising COMPOUND 1 or COMPOUND 2 is prepared as acontrolled release tablet or capsule which includes a drug corecomprising the pharmaceutical composition and optional excipients.Optionally, a “seal coat” or “shell” is applied. In certain embodiments,a formulation provided herein comprising COMPOUND 1 or COMPOUND 2provided herein is a controlled release tablet or capsule, whichcomprises a therapeutically effective amount of COMPOUND 1 or COMPOUND2, a drug release controlling component that controls the release ofCOMPOUND 1 or COMPOUND 2 substantially in the stomach upon oraladministration, and optionally, one or more excipients.

Particular embodiments provide a drug release controlling component thatis a polymer matrix, which swells upon exposure to gastric fluid toeffect the gastric retention of the formulation and the sustainedrelease of COMPOUND 1 or COMPOUND 2 from the polymer matrixsubstantially in the stomach. In certain embodiments, such formulationsmay be prepared by incorporating COMPOUND 1 or COMPOUND 2 into asuitable polymeric matrix during formulation. Examples of suchformulations are known in the art. See, e.g., Shell et al., U.S. PatentPublication No. 2002/0051820 (application Ser. No. 09/990,061); Shell etal., U.S. Patent Publication No. 2003/0039688 (application Ser. No.10/045,823); Gusler et al., U.S. Patent Publication No. 2003/0104053(application Ser. No. 10/029,134), each of which is incorporated hereinby reference in its entirety.

In certain embodiments, the drug release controlling component maycomprise a shell surrounding the drug-containing core, wherein the shellreleases COMPOUND 1 or COMPOUND 2 from the core by, e.g., permittingdiffusion of COMPOUND 1 or COMPOUND 2 from the core and promotinggastric retention of the formulation by swelling upon exposure togastric fluids to a size that is retained in the stomach. In certainembodiments, such formulations may be prepared by first compressing amixture of COMPOUND 1 or COMPOUND 2 and one or more excipients to form adrug core, and compressing another powdered mixture over the drug coreto form the shell, or enclosing the drug core with a capsule shell madeof suitable materials. Examples of such formulations are known in theart. See, e.g., Berner et al., U.S. Patent Publication No. 2003/0104062application Ser. No. 10/213,823), incorporated herein by reference inits entirety.

In certain embodiments, the pharmaceutical formulations provided hereincontain COMPOUND 1 or COMPOUND 2 and, optionally, one or more excipientsto form a “drug core.” Optional excipients include, e.g., diluents(bulking agents), lubricants, disintegrants, fillers, stabilizers,surfactants, preservatives, coloring agents, flavoring agents, bindingagents, excipient supports, glidants, permeation enhancement excipients,plasticizers and the like, e.g., as known in the art. It will beunderstood by those in the art that some substances serve more than onepurpose in a pharmaceutical composition. For instance, some substancesare binders that help hold a tablet together after compression, yet arealso disintegrants that help break the tablet apart once it reaches thetarget delivery site. Selection of excipients and amounts to use may bereadily determined by the formulation scientist based upon experienceand consideration of standard procedures and reference works availablein the art.

In certain embodiments, formulations provided herein comprise one ormore binders. Binders may be used, e.g., to impart cohesive qualities toa tablet, and thus ensure that the tablet remains intact aftercompression. Suitable binders include, but are not limited to, starch(including corn starch and pregelatinized starch), gelatin, sugars(including sucrose, glucose, dextrose and lactose), polyethylene glycol,propylene glycol, waxes, and natural and synthetic gums, e.g., acaciasodium alginate, polyvinylpyrrolidone, cellulosic polymers (includinghydroxypropyl cellulose, hydroxypropylmethylcellulose, methyl cellulose,ethyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose and thelike), veegum, carbomer (e.g., carbopol), sodium, dextrin, guar gum,hydrogenated vegetable oil, magnesium aluminum silicate, maltodextrin,polymethacrylates, povidone (e.g., KOLLIDON, PLASDONE), microcrystallinecellulose, among others. Binding agents also include, e.g., acacia,agar, alginic acid, cabomers, carrageenan, cellulose acetate phthalate,ceratonia, chitosan, confectioner's sugar, copovidone, dextrates,dextrin, dextrose, ethylcellulose, gelatin, glyceryl behenate, guar gum,hydroxyethyl cellulose, hydroxyethylmethyl cellulose, hydroxypropylcellulose, hydroxypropyl starch, hypromellose, inulin, lactose,magnesium aluminum silicate, maltodextrin, maltose, methylcellulose,poloxamer, polycarbophil, polydextrose, polyethylene oxide,polymethylacrylates, povidone, sodium alginate, sodiumcarboxymethylcellulose, starch, pregelatinized starch, stearic acid,sucrose, and zein. The binding agent can be, relative to the drug core,in the amount of about 2% w/w of the drug core; about 4% w/w of the drugcore, about 6% w/w of the drug core, about 8% w/w of the drug core,about 10% w/w of the drug core, about 12% w/w of the drug core, about14% w/w of the drug core, about 16% w/w of the drug core, about 18% w/wof the drug core, about 20% w/w of the drug core, about 22% w/w of thedrug core, about 24% w/w of the drug core, about 26% w/w of the drugcore, about 28% w/w of the drug core, about 30% w/w of the drug core,about 32% w/w of the drug core, about 34% w/w of the drug core, about36% w/w of the drug core, about 38% w/w of the drug core, about 40% w/wof the drug core, about 42% w/w of the drug core, about 44% w/w of thedrug core, about 46% w/w of the drug core, about 48% w/w of the drugcore, about 50% w/w of the drug core, about 52% w/w of the drug core,about 54% w/w of the drug core, about 56% w/w of the drug core, about58% w/w of the drug core, about 60% w/w of the drug core, about 62% w/wof the drug core, about 64% w/w of the drug core, about 66% w/w of thedrug core; about 68% w/w of the drug core, about 70% w/w of the drugcore, about 72% w/w of the drug core, about 74% w/w of the drug core,about 76% w/w of the drug core, about 78% w/w of the drug core, about80% w/w of the drug core, about 82% w/w of the drug core, about 84% w/wof the drug core, about 86% w/w of the drug core, about 88% w/w of thedrug core, about 90% w/w of the drug core, about 92% w/w of the drugcore, about 94% w/w of the drug core, about 96% w/w of the drug core,about 98% w/w of the drug core, or more, if determined to beappropriate. In certain embodiments, a suitable amount of a particularbinder is determined by one of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore diluents. Diluents may be used, e.g., to increase bulk so that apractical size tablet is ultimately provided. Suitable diluents includedicalcium phosphate, calcium sulfate, lactose, cellulose, kaolin,mannitol, sodium chloride, dry starch, microcrystalline cellulose (e.g.,AVICEL), microfine cellulose, pregelitinized starch, calcium carbonate,calcium sulfate, sugar, dextrates, dextrin, dextrose, dibasic calciumphosphate dihydrate, tribasic calcium phosphate, kaolin, magnesiumcarbonate, magnesium oxide, maltodextrin, mannitol, polymethacrylates(e.g., EUDRAGIT), potassium chloride, sodium chloride, sorbitol andtalc, among others. Diluents also include, e.g., ammonium alginate,calcium carbonate, calcium phosphate, calcium sulfate, celluloseacetate, compressible sugar, confectioner's sugar, dextrates, dextrin,dextrose, erythritol, ethylcellulose, fructose, fumaric acid, glycerylpalmitostearate, isomalt, kaolin, lacitol, lactose, mannitol, magnesiumcarbonate, magnesium oxide, maltodextrin, maltose, medium-chaintriglycerides, microcrystalline cellulose, microcrystalline silicifiedcellulose, powered cellulose, polydextrose, polymethylacrylates,simethicone, sodium alginate, sodium chloride, sorbitol, starch,pregelatinized starch, sucrose, sulfobutylether-β-cyclodextrin, talc,tragacanth, trehalose, and xylitol. Diluents may be used in amountscalculated to obtain a desired volume for a tablet or capsule; incertain embodiments, a diluent is used in an amount of about 5% or more,about 10% or more, about 15% or more, about 20% or more, about 22% ormore, about 24% or more, about 26% or more, about 28% or more, about 30%or more, about 32% or more, about 34% or more, about 36% or more, about38% or more, about 40% or more, about 42% or more, about 44% or more,about 46% or more, about 48% or more, about 50% or more, about 52% ormore, about 54% or more, about 56% or more, about 58% or more, about 60%or more, about 62% or more, about 64% or more, about 68% or more, about70% ore more, about 72% or more, about 74% or more, about 76% or more,about 78% or more, about 80% or more, about 85% or more, about 90% ormore, or about 95% or more, weight/weight, of a drug core; between about10% and about 90% w/w of the drug core; between about 20% and about 80%w/w of the drug core; between about 30% and about 70% w/w of the drugcore; between about 40% and about 60% w/w of the drug core. In certainembodiments, a suitable amount of a particular diluent is determined byone of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore lubricants. Lubricants may be used, e.g., to facilitate tabletmanufacture; examples of suitable lubricants include, for example,vegetable oils such as peanut oil, cottonseed oil, sesame oil, oliveoil, corn oil, and oil of theobroma, glycerin, magnesium stearate,calcium stearate, and stearic acid. In certain embodiments, stearates,if present, represent no more than approximately 2 weight % of thedrug-containing core. Further examples of lubricants include, e.g.,calcium stearate, glycerin monostearate, glyceryl behenate, glycerylpalmitostearate, magnesium lauryl sulfate, magnesium stearate, myristicacid, palmitic acid, poloxamer, polyethylene glycol, potassium benzoate,sodium benzoate, sodium chloride, sodium lauryl sulfate, sodium stearylfumarate, stearic acid, talc, and zinc stearate. In particularembodiments, the lubricant is magnesium stearate. In certainembodiments, the lubricant is present, relative to the drug core, in anamount of about 0.2% w/w of the drug core, about 0.4% w/w of the drugcore, about 0.6% w/w of the drug core, about 0.8% w/w of the drug core,about 1.0% w/w of the drug core, about 1.2% w/w of the drug core, about1.4% w/w of the drug core, about 1.6% w/w of the drug core, about 1.8%w/w of the drug core, about 2.0% w/w of the drug core, about 2.2% w/w ofthe drug core, about 2.4% w/w of the drug core, about 2.6% w/w of thedrug core, about 2.8% w/w of the drug core, about 3.0% w/w of the drugcore, about 3.5% w/w of the drug core, about 4% w/w of the drug core,about 4.5% w/w of the drug core, about 5% w/w of the drug core, about 6%w/w of the drug core, about 7% w/w of the drug core, about 8% w/w of thedrug core, about 10% w/w of the drug core, about 12% w/w of the drugcore, about 14% w/w of the drug core, about 16% w/w of the drug core,about 18% w/w of the drug core, about 20% w/w of the drug core, about25% w/w of the drug core, about 30% w/w of the drug core, about 35% w/wof the drug core, about 40% w/w of the drug core, between about 0.2% andabout 10% w/w of the drug core, between about 0.5% and about 5% w/w ofthe drug core, or between about 1% and about 3% w/w of the drug core. Incertain embodiments, a suitable amount of a particular lubricant isdetermined by one of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore disintegrants. Disintegrants may be used, e.g., to facilitatedisintegration of the tablet, and may be, e.g., starches, clays,celluloses, algins, gums or crosslinked polymers. Disintegrants alsoinclude, e.g., alginic acid, carboxymethylcellulose calcium,carboxymethylcellulose sodium (e.g., AC-DI-SOL, PRIMELLOSE), colloidalsilicon dioxide, croscarmellose sodium, crospovidone (e.g., KOLLIDON,POLYPLASDONE), guar gum, magnesium aluminum silicate, methyl cellulose,microcrystalline cellulose, polacrilin potassium, powdered cellulose,pregelatinized starch, sodium alginate, sodium starch glycolate (e.g.,EXPLOTAB) and starch. Additional disintegrants include, e.g., calciumalginate, chitosan, sodium docusate, hydroxypropyl cellulose, andpovidone. In certain embodiments, the disintegrant is, relative to thedrug core, present in the amount of about 1% w/w of the drug core, about2% w/w of the drug core, about 3% w/w of the drug core, about 4% w/w ofthe drug core, about 5% w/w of the drug core, about 6% w/w of the drugcore, about 7% w/w of the drug core, about 8% w/w of the drug core,about 9% w/w of the drug core, about 10% w/w of the drug core, about 12%w/w of the drug core, about 14% w/w of the drug core, about 16% w/w ofthe drug core, about 18% w/w of the drug core, about 20% w/w of the drugcore, about 22% w/w of the drug core, about 24% w/w of the drug core,about 26% w/w of the drug core, about 28% w/w of the drug core, about30% w/w of the drug core, about 32% w/w of the drug core, greater thanabout 32% w/w of the drug core, between about 1% and about 10% w/w ofthe drug core, between about 2% and about 8% w/w of the drug core,between about 3% and about 7% w/w of the drug core, or between about 4%and about 6% w/w of the drug core. In certain embodiments, a suitableamount of a particular disintegrant is determined by one of ordinaryskill in the art.

In certain embodiments, formulations provided herein comprise one ormore stabilizers. Stabilizers (also called absorption enhancers) may beused, e.g., to inhibit or retard drug decomposition reactions thatinclude, by way of example, oxidative reactions. Stabilizing agentsinclude, e.g., d-Alpha-tocopheryl polyethylene glycol 1000 succinate(Vitamin E TPGS), acacia, albumin, alginic acid, aluminum stearate,ammonium alginate, ascorbic acid, ascorbyl palmitate, bentonite,butylated hydroxytoluene, calcium alginate, calcium stearate, calciumcarboxymethylcellulose, carrageenan, ceratonia, colloidal silicondioxide, cyclodextrins, diethanolamine, edetates, ethylcellulose,ethyleneglycol palmitostearate, glycerin monostearate, guar gum,hydroxypropyl cellulose, hypromellose, invert sugar, lecithin, magnesiumaluminum silicate, monoethanolamine, pectin, poloxamer, polyvinylalcohol, potassium alginate, potassium polacrilin, povidone, propylgallate, propylene glycol, propylene glycol alginate, raffinose, sodiumacetate, sodium alginate, sodium borate, sodium carboxymethyl cellulose,sodium stearyl fumarate, sorbitol, stearyl alcohol,sufobutyl-b-cyclodextrin, trehalose, white wax, xanthan gum, xylitol,yellow wax, and zinc acetate. In certain embodiments, the stabilizer is,relative to the drug core, present in the amount of about 1% w/w of thedrug core, about 2% w/w of the drug core, about 3% w/w of the drug core,about 4% w/w of the drug core, about 5% w/w of the drug core, about 6%w/w of the drug core, about 7% w/w of the drug core, about 8% w/w of thedrug core, about 9% w/w of the drug core, about 10% w/w of the drugcore, about 12% w/w of the drug core, about 14% w/w of the drug core,about 16% w/w of the drug core, about 18% w/w of the drug core, about20% w/w of the drug core, about 22% w/w of the drug core, about 24% w/wof the drug core, about 26% w/w of the drug core, about 28% w/w of thedrug core, about 30% w/w of the drug core, about 32% w/w of the drugcore, between about 1% and about 10% w/w of the drug core, between about2% and about 8% w/w of the drug core, between about 3% and about 7% w/wof the drug core, or between about 4% and about 6% w/w of the drug core.In certain embodiments, a suitable amount of a particular stabilizer isdetermined by one of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore glidants. Glidants may be used, e.g., to improve the flowproperties of a powder composition or granulate or to improve theaccuracy of dosing. Excipients that may function as glidants include,e.g., colloidal silicon dioxide, magnesium trisilicate, powderedcellulose, starch, tribasic calcium phosphate, calcium silicate,powdered cellulose, colloidal silicon dioxide, magnesium silicate,magnesium trisilicate, silicon dioxide, starch, tribasic calciumphosphate, and talc. In certain embodiments, the glidant is, relative tothe drug core, present in the amount of less than about 1% w/w of thedrug core, about 1% w/w of the drug core, about 2% w/w of the drug core,about 3% w/w of the drug core, about 4% w/w of the drug core, about 5%w/w of the drug core, about 6% w/w of the drug core, about 7% w/w of thedrug core, about 8% w/w of the drug core, about 9% w/w of the drug core,about 10% w/w of the drug core, about 12% w/w of the drug core, about14% w/w of the drug core, about 16% w/w of the drug core, about 18% w/wof the drug core, about 20% w/w of the drug core, about 22% w/w of thedrug core, about 24% w/w of the drug core, about 26% w/w of the drugcore, about 28% w/w of the drug core, about 30% w/w of the drug core,about 32% w/w of the drug core, between about 1% and about 10% w/w ofthe drug core, between about 2% and about 8% w/w of the drug core,between about 3% and about 7% w/w of the drug core, or between about 4%and about 6% w/w of the drug core. In certain embodiments, a suitableamount of a particular glidant is determined by one of ordinary skill inthe art.

In one embodiment, the pharmaceutical compositions provided herein maybe administered orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or via an implanted reservoir,preferably by oral administration or administration by injection. In oneembodiment, the pharmaceutical compositions may contain any conventionalnon-toxic pharmaceutically-acceptable carriers, adjuvants or vehicles.In some cases, the pH of the formulation may be adjusted withpharmaceutically acceptable acids, bases or buffers to enhance thestability of the formulated compound or its delivery form. The termparenteral as used herein includes subcutaneous, intracutaneous,intravenous, intramuscular, intraarticular, intraarterial,intrasynovial, intrasternal, intrathecal, intralesional and intracranialinjection or infusion techniques.

In one embodiment, the pharmaceutical compositions provided herein maybe in the form of a sterile injectable preparation, for example, as asterile injectable aqueous or oleaginous suspension. This suspension maybe formulated according to techniques known in the art using suitabledispersing or wetting agents (such as, for example, Tween 80) andsuspending agents. The sterile injectable preparation may also be asterile injectable solution or suspension in a non-toxic parenterallyacceptable diluent or solvent, for example, as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are mannitol, water, Ringer's solution and isotonic sodiumchloride solution. In addition, sterile, fixed oils are conventionallyemployed as a solvent or suspending medium. For this purpose, any blandfixed oil may be employed including synthetic mono- or diglycerides.Fatty acids, such as oleic acid and its glyceride derivatives are usefulin the preparation of injectables, as are naturalpharmaceutically-acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions may also contain a long-chain alcohol diluent or dispersant,or carboxymethyl cellulose or similar dispersing agents which arecommonly used in the formulation of pharmaceutically acceptable dosageforms such as emulsions and or suspensions. Other commonly usedsurfactants such as Tweens or Spans and/or other similar emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

In one embodiment, the pharmaceutical compositions provided herein mayalso be administered in the form of suppositories for rectaladministration. These compositions can be prepared by mixing a compoundof one aspect of this invention with a suitable non-irritating excipientwhich is solid at room temperature but liquid at the rectal temperatureand therefore will melt in the rectum to release the active components.Such materials include, but are not limited to, cocoa butter, beeswaxand polyethylene glycols.

Topical administration of the pharmaceutical compositions providedherein is useful when the desired treatment involves areas or organsreadily accessible by topical application. For application topically tothe skin, the pharmaceutical composition should be formulated with asuitable ointment containing the active components suspended ordissolved in a carrier. Carriers for topical administration of thecompounds of one aspect of this invention include, but are not limitedto, mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier with suitable emulsifying agents. Suitablecarriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water. The pharmaceuticalcompositions provided herein may also be topically applied to the lowerintestinal tract by rectal suppository formulation or in a suitableenema formulation. Topically-transdermal patches are also includedherein.

In one embodiment, the pharmaceutical compositions provided herein maybe administered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other solubilizing or dispersingagents known in the art.

In one embodiment, the compositions provided herein can, for example, beadministered by injection, intravenously, intraarterially, subdermally,intraperitoneally, intramuscularly, or subcutaneously; or orally,buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.5 toabout 100 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. In one embodiment, the pharmaceuticalcompositions are administered from about 1 to about 6 times per day oralternatively, as a continuous infusion. Such administration can be usedas a chronic or acute therapy. The amount of active ingredient that maybe combined with the carrier materials to produce a single dosage formvaries depending upon the host treated and the particular mode ofadministration. A typical preparation contains from about 5% to about95% active compound (w/w). Alternatively, such preparations contain fromabout 20% to about 80% active compound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular subject depends upon avariety of factors, including the activity of the specific compoundemployed, the age, body weight, general health status, sex, diet, timeof administration, rate of excretion, drug combination, the severity andcourse of the disease, condition or symptoms, the subject's dispositionto the disease, condition or symptoms, and the judgment of the treatingphysician.

Upon improvement of a subject's condition, a maintenance dose of acompound, composition or combination provided herein may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Subjects may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

Solid Dispersions of COMPOUND 2

In certain embodiment, COMPOUND 2 is administered in compositions,comprising COMPOUND 2, and one or more polymer(s) as part of a soliddispersion (e.g., an amorphous solid dispersion). In some embodiments,the solid dispersion comprises COMPOUND 2, and one or more polymer(s).In some embodiments, the solid dispersion comprises COMPOUND 2, one ormore polymer(s), and one or more surfactant(s). In some embodiments, thesolid dispersion comprises COMPOUND 2, and one polymer. In someembodiments, the solid dispersion comprises COMPOUND 2, one polymer, anda surfactant.

In certain embodiment, the solid dispersions provided herein, comprisingCOMPOUND 2, enhance the solubility of COMPOUND 2 relative to a neatcrystalline form of COMPOUND 2 (e.g., Form 1 or Form 2), and thusprovide improved exposure upon oral dosing of the solid dispersion to asubject. In one embodiment, the solid dispersion comprises COMPOUND 2,one or more polymer(s), and optionally one or more solubility enhancingsurfactant.

For example, the aqueous solubility of Form 1 is about 0.025 mg/mL toabout 0.035 mg/mL and the aqueous solubility of Form 2 is about 0.008mg/mL to about 0.010 mg/mL.

Form 2 has a solubility of about 0.018 mg/mL in fasted state simulatedintestinal fluid (FASSIF) at a pH of 6.1 at 4 hours. In comparison,amorphous spray-dried dispersions have a solubility of about 0.05 mg/mLto about 0.50 mg/mL in FASSIF at 3 hours.

In some embodiments, the solid dispersion exhibits at least about 20%,at least about 30%, at least about 40%, at least about 50%, at leastabout 60%, at least about 70%, at least about 80%, or at least about 90%higher exposure of COMPOUND 2, when administered to a subject ascompared to administration of in-situ amorphous COMPOUND 2. In someembodiments, the solid dispersion exhibits at least about 20%, at leastabout 30%, at least about 40%, at least about 50%, at least about 60%,at least about 70%, at least about 80%, or at least about 90% higherexposure of COMPOUND 2, when administered to a subject as compared toadministration of neat crystalline COMPOUND 2.

In rat and monkey pharmacokinetics studies, modest exposure improvementis observed upon administration of solid dispersion oral dosage forms ascompared to in-situ amorphous dosing shows. For example, a soliddispersion containing 50% w/w COMPOUND 2 and 50% w/w Polyvinyl AcetatePhthalate (PVAP) has approximately two-fold higher exposure as comparedto in-situ amorphous COMPOUND 2 in male Sprague Dawley rats. There is nosignificant difference in exposure between a solid dispersion containing70% w/w COMPOUND 2 and 30% w/w oral dosage form as compared to in-situamorphous COMPOUND 2. In male cynomolgus monkeys, the exposure of asolid dispersion containing 50% w/w COMPOUND 2 and 50% w/whydroxypropylmethylcellulose acetate succinate, also known ashpromellose acetate succinate, (HPMCAS) shows no significant differenceas compared to the in-situ amorphous COMPOUND 2. Similarly, a soliddispersion containing 50% w/w COMPOUND 2 and 50% w/whydroxypropylmethylcellulose also known as hypromellose phthalate(HPMC-Phthalate) shows no significant difference as compared to thein-situ amorphous COMPOUND 2. While in-situ amorphous therapeuticcompounds are commonly used for dosing in animal studies, they are notsuitable dosage forms for dosing in humans.

As described in the rat pharmacokinetics study of Example 4, COMPOUND 2exposure is improved when solid dispersion dosage forms are administeredas compared to neat crystalline COMPOUND 2 Form 2.

In some embodiments, at least a portion of COMPOUND 2, in the soliddispersion is in the amorphous state (e.g., at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, at least about 80%, at least about 85%, at leastabout 90%, at least about 95%, at least about 98%, or at least about99%). In other embodiments, the solid dispersion is substantially freeof crystalline COMPOUND 2.

In some embodiments, the composition is an amorphous solid (e.g. spraydried) dispersion comprising COMPOUND 2, and a polymer. The amorphoussolid dispersion can include, e.g., less than about 30%, less than about20%, less than about 15%, less than about 10%, less than about 5%, lessthan about 4%, less than about 3%, less than about 2%, or less thanabout 1% of crystalline COMPOUND 2, e.g., be substantially free ofcrystalline COMPOUND 2.

In one embodiment, the solid dispersion exhibits a predetermined levelof physical and/or chemical stability. E.g., the solid dispersionretains about 50%, about 60%, about 70%, about 80%, about 90%, about95%, about 98%, or about 99%, of amorphous COMPOUND 2, when stored at25° C. in a closed water tight container, e.g., an amber glass vial,high density polyethylene (HDPE) container or double polyethylene bagswith twisted nylon tie placed in an HDPE container with desiccant.

In some embodiments, the polymer increases the chemical or physicalstability (e.g., as measured by a Modulated Differential Scanningcalorimeter) of COMPOUND 2, when stored (e.g., at 2-8° C., e.g. 4° C. orat room temperature) by at least about 10% (e.g., by at least about 20%,by at least about 30%, by at least about 40%, by at least about 50%, byat least about 60%, by at least about 70%, by at least about 80%, or byat least about 90%) compared to amorphous COMPOUND 2, without being inthe presence of the polymer.

A solid dispersion generally exhibits a glass transition temperature,where the dispersion makes a transition from a glassy solid to a rubberycomposition. In general, the higher the glass transition temperature,the greater the physical stability of the dispersion. The existence of aglass transition temperature generally indicates that at least a largeportion of the composition (e.g., dispersion) is in an amorphous state.The glass transition temperature (Tg) of a solid dispersion suitable forpharmaceutical applications is generally at least about 50° C. In someembodiments, higher temperatures are preferred. Therefore, in someembodiments, a solid dispersion disclosed herein has a Tg of at leastabout 100° C. (e.g., at least about 100° C., at least about 105° C., atleast about 110° C., at least about 115° C., at least about 120° C., atleast about 125° C., at least about 130° C., at least about 135° C., atleast about 140° C., at least about 150° C., at least about 160° C., atleast about 170° C., at least about 175° C., at least about 180° C., orat least about 190° C.). In some embodiments, the Tg is up to about 200°C. In some embodiments, the Tg is up to about 130° C. (e.g., at leastabout 110° C., at least about 111° C., at least about 112° C., at leastabout 113° C., at least about 114° C., at least about 115° C., at leastabout 116° C., at least about 117° C., at least about 118° C., at leastabout 119° C., at least about 120° C., at least about 121° C., at leastabout 122° C., at least about 123° C., at least about 124° C., at leastabout 125° C., at least about 1216° C., at least about 127° C., at leastabout 128° C., at least about 129° C., or at least about 130° C.).Unless otherwise noted, the glass transition temperatures disclosedherein are measured under dry conditions.

In some embodiments the solid dispersion has a higher glass transitiontemperature than the glass transition temperature of amorphous COMPOUND2, without being in the presence of the polymer(s). In some embodiments,the solid dispersion has a relaxation rate that is lower than therelaxation rate of amorphous COMPOUND 2, without being in the presenceof the polymer(s).

Examples of polymers in the solid dispersion include cellulosederivatives (e.g., hydroxypropylmethylcellulose also known ashypromellose, (HPMC), hydroxypropylmethylcellulose phthalate, also knownas hypromellose phthalate (HPMCP), hydroxypropylmethylcellulose acetatesuccinate, also known as hpromellose acetate succinate, (HPMCAS),hydroxypropylcellulose (HPC)), ethylcellulose, or cellulose acetatephthalate; polyvinylpyrrolidones (PVP); polyethylene glycols (PEG);polyvinyl alcohols (PVA); polyvinyl esters, such as Polyvinyl AcetatePhthalate (PVAP); acrylates, such as polymethacrylate (e.g., Eudragit®E); cyclodextrins (e.g., .beta.-cyclodextrin); Poly (D, L-lactide)(PLA), Poly (D,L-lactide, co-glycolide acid (PLGA); and copolymers andderivatives thereof, including for example polyvinylpyrollidone-vinylacetate (PVP-VA), Polyvinyl caprolactam-polyvinyl, andacetate-polyethyleneglycol copolymer, Methylacrylate/methacrylic acidcopolymer; Soluplus; Copovidone; and mixtures thereof.

In some embodiments, the solid dispersion includes one water-solublepolymer. In some embodiments, the solid dispersion includes onepartially water-soluble polymer. In some embodiments, the polymer is acellulose polymer.

In some embodiments, the polymer is HPMCAS (e.g., HPMCAS of differentgrades: HPMCAS-M, HPMCAS-MG or HPMCAS-HG). In some embodiments, thepolymer is PVAP. In some embodiments, the polymer is HPMC (e.g., HPMC ofdifferent grades: HMPC60SH50, HPMCE50 or HPMCE15). In some embodiments,the polymer is HPMCP (e.g., HPMCP of different grades: e.g.,HMPCP-HP55).

In some embodiments, the polymer is a pH-dependent enteric polymer. SuchpH-dependent enteric polymers include, but are not limited to, cellulosederivatives (e.g., cellulose acetate phthalate (CAP)), HPMCP, HPMCAS,carboxymethylcellulose (CMC) or a salt thereof (e.g., a sodium salt suchas (CMC-Na)); cellulose acetate trimellitate (CAT),hydroxypropylcellulose acetate phthalate (HPCAP),hydroxypropylmethyl-cellulose acetate phthalate (HPMCAP), andmethylcellulose acetate phthalate (MCAP), polymethacrylates (e.g.,Eudragit S), or mixtures thereof.

In some embodiments, the polymer is hydroxypropylmethylcellulose acetatesuccinate, also known as hypromellose acetate succinate, (HPMCAS), e.g.,HMPCAS-HG.

In another embodiment, the polymer(s) is an insoluble cross-linkedpolymer, for example a polyvinylpyrrolidone (e.g., Crospovidone). Inanother embodiment, the polymer(s) is polyvinylpyrrolidone (PVP).

In some embodiments, the one or more polymer(s) is present in the soliddispersion in an amount of between about 10% w/w and 90% w/w (e.g.,between about 20% w/w and about 80% w/w; between about 30% w/w and about70% w/w; between about 40% w/w and about 60% w/w; or between about 15%w/w and about 35% w/w). In some embodiments, the polymer(s) is presentin the solid dispersion in an amount of from about 10% w/w to about 80%w/w, for example from about 30% w/w to about 75% w/w, or from about 40%w/w to about 65% w/w, or from about 45% w/w to about 55% w/w, forexample, about 46% w/w, about 47% w/w, about 48% w/w, about 49% w/w,about 50% w/w, about 51% w/w, about 52% w/w, about 53% w/w, or about 54%w/w. In some embodiments, the polymer(s) is present in the soliddispersion in an amount of about 48% w/w, about 48.5% w/w, about 49%w/w, about 49.5% w/w, about 50% w/w, about 50.5% w/w, about 51% w/w,about 51.5% w/w, about 52% w/w, or about 52.5% w/w.

In some embodiments, the polymer(s) is present in the solid dispersionin an amount of from about 30% w/w to about 70% w/w. In someembodiments, the polymer(s) is present in the solid dispersion in anamount of from about 35% w/w to about 65% w/w. In some embodiments, thepolymer(s) is present in the solid dispersion in an amount of from about40% w/w to about 60% w/w. In some embodiments, the polymer(s) is presentin the solid dispersion in an amount of from about 45% w/w to about 55%w/w. In some embodiments, the polymer(s) is present in the soliddispersion in an amount of about 50% w/w.

In some embodiments, COMPOUND 2, is present in the solid dispersion inan amount of from about 10% w/w and 90% w/w (e.g., between about 20% w/wand about 80% w/w; between about 30% w/w and about 70% w/w; betweenabout 40% w/w and about 60% w/w; or between about 15% w/w and about 35%w/w). In some embodiments, COMPOUND 2, is present in the soliddispersion in an amount of from about 10% w/w to about 80% w/w, forexample from about 30% w/w to about 75% w/w, or from about 40% w/w toabout 65% w/w, or from about 45% w/w to about 55% w/w, for example,about 46% w/w, about 47% w/w, about 48% w/w, about 49% w/w, about 50%w/w, about 51% w/w, about 52% w/w, about 53% w/w, or about 54% w/w. Insome embodiments, COMPOUND 2, is present in the solid dispersion in anamount of about 48% w/w, about 48.5% w/w, about 49% w/w, about 49.5%w/w, about 50% w/w, about 50.5% w/w, about 51% w/w, about 51.5% w/w,about 52% w/w, or about 52.5% w/w.

In some embodiments, COMPOUND 2, is present in the solid dispersion inan amount of from about 30% w/w to about 70% w/w. In some embodiments,COMPOUND 2, is present in the solid dispersion in an amount of fromabout 35% w/w to about 65% w/w. In some embodiments, COMPOUND 2, ispresent in the solid dispersion in an amount of from about 40% w/w toabout 60% w/w. In some embodiments, COMPOUND 2, is present in the soliddispersion in an amount of from about 45% w/w to about 55% w/w. In someembodiments, COMPOUND 2, is present in the solid dispersion in an amountof about 50% w/w.

In another embodiment, the solid dispersion includes about 20% w/w toabout 80% w/w COMPOUND 2, and about 20% w/w to about 80% of polymer(s).In another embodiment, the solid dispersion includes about 25% w/w toabout 75% w/w COMPOUND 2, and about 25% w/w to about 75% of polymer(s).In another embodiment, the solid dispersion includes about 30% w/w toabout 70% w/w COMPOUND 2, and about 30% w/w to about 70% of polymer(s).In another embodiment, the solid dispersion includes about 35% w/w toabout 65% w/w COMPOUND 2, and about 35% w/w to about 65% of polymer(s).In another embodiment, the solid dispersion includes about 40% w/w toabout 60% w/w COMPOUND 2, and about 40% w/w to about 60% of polymer(s).In another embodiment, the solid dispersion includes about 45% w/w toabout 55% w/w COMPOUND 2, and about 45% w/w to about 55% of polymer(s).In another embodiment, the solid dispersion includes about 50% w/wCOMPOUND 2, and about 50% w/w of polymer(s).

In another embodiment, the solid dispersion includes about 45% w/w toabout 55% w/w COMPOUND 2, and about 45% w/w to about 55% w/w HPMCAS(e.g., HPMCAS-MG or HPMCAS-HG, or other grades such as LF, MF, HF, orLG) or PVAP. In another embodiment, the solid dispersion includes about50% w/w COMPOUND 2, and about 50% w/w of HPMCAS.

In some embodiments, the solid dispersion also includes a surfactant orinert pharmaceutically acceptable substance. Examples of surfactants inthe solid dispersion include sodium lauryl sulfate (SLS), vitamin E or aderivative thereof (e.g., vitamin E TPGS), Docusate Sodium, sodiumdodecyl sulfate, polysorbates (such as Tween 20 and Tween 80),poloxamers (such as Poloxamer 335 and Poloxamer 407), glycerylmonooleate, Span 65, Span 25, Capryol 90, pluronic copolymers (e.g.,Pluronic F108, Pluronic P-123), and mixtures thereof. In someembodiments, the surfactant is SLS. In some embodiments, the surfactantis vitamin E or a derivative thereof (e.g., vitamin E TPGS).

In some embodiments, the surfactant is present in the solid dispersionin an amount of from about 0.1% w/w to about 10% w/w, for example fromabout 0.5% w/w to about 2% w/w, or from about 1% w/w to about 3% w/w,from about 1% w/w to about 4% w/w, or from about 1% w/w to about 5% w/w.In some embodiments, the surfactant is present in the solid dispersionin an amount of about 0.1% w/w, about 0.2% w/w, about 0.3% w/w, about0.4% w/w, about 0.5% w/w, about 0.6% w/w, about 0.7% w/w, about 0.8%w/w, about 0.9% w/w, or about 1% w/w. In some embodiments, thesurfactant is present in the solid dispersion in an amount of about 0.5%w/w, about 1% w/w, about 1.5% w/w, about 2% w/w, about 2.5% w/w, about3% w/w, about 3.5% w/w, about 4% w/w, about 4.5% w/w, or about 5% w/w.

Processes for Preparing Solid Dispersions

In some embodiments, the solid dispersion may be prepared according to aprocess described herein. In general, methods that could be used includethose that involve rapid removal of solvent or solvent mixture from amixture or cooling a molten sample. Such methods include, but are notlimited to, rotational evaporation, freeze-drying (i.e.,lyophilization), vacuum drying, melt congealing, and melt extrusion. Oneembodiment of this disclosure involves solid dispersion obtained byspray-drying. In one embodiment, the product obtained by spray drying isdried to remove the solvent or solvent mixture.

Preparations disclosed herein, e.g., a pharmaceutical composition, canbe obtained by spray-drying a mixture comprising COMPOUND 2, one or morepolymer(s), and an appropriate solvent or solvent mixture. Spray dryinginvolves atomization of a liquid mixture containing, e.g., a solid and asolvent or solvent mixture, and removal of the solvent or solventmixture. The solvent or solvent mixture can also contain a nonvolatilesolvent, such as glacial acetic acid. Atomization may be done, forexample, through a two-fluid or pressure or electrosonic nozzle or on arotating disk.

Spray drying converts a liquid feed to a dried particulate form. Spraydrying generally involves the atomization of a liquid feed solution intoa spray of droplets and contacting the droplets with hot air or gas in adrying chamber. The sprays are generally produced by either rotary(wheel) or nozzle atomizers. Evaporation of moisture from the dropletsand formation of dry particles proceed under controlled temperature andairflow conditions.

Optionally, a secondary drying process such as fluidized bed drying orvacuum drying, may be used to reduce residual solvents (and otheradditives, such as glacial acetic acid) to pharmaceutically acceptablelevels. Typically, spray-drying involves contacting a highly dispersedliquid suspension or solution (e.g., atomized solution), and asufficient volume of hot air or gas (e.g., nitrogen, e.g., purenitrogen) to produce evaporation and drying of the liquid droplets. Thepreparation to be spray dried can be any solution, coarse suspension,slurry, colloidal dispersion, or paste that may be atomized using theselected spray-drying apparatus. In a standard procedure, thepreparation is sprayed into a current of warm filtered air (or into gas,e.g., nitrogen) that evaporates the solvent and conveys the driedproduct to a collector (e.g., a cyclone). The spent air or gas is thenexhausted with the solvent (or solvent mixture including any additivessuch as glacial acetic acid), (e.g., then filtered) or alternatively thespent air or gas is sent to a condenser to capture and potentiallyrecycle the solvent or solvent mixture. For example, if a gas (e.g.,nitrogen) is used, the gas is then optionally recycled, heated again andreturned to the unit in a closed loop system. Commercially availabletypes of apparatus may be used to conduct the spray-drying. For example,commercial spray dryers are manufactured by Buchi Ltd. and Niro (e.g.,the PSD line of spray driers manufactured by Niro).

Spray-drying typically employs solids loads of material from about 1% toabout 30% or up to about 50% (i.e., therapeutically active compound plusand excipients), preferably at least about 10%. In some embodiments,solids loads of less than 10% may result in poor yields and unacceptablylong run-times. In general, the upper limit of solids loads is governedby the viscosity of (e.g., the ability to pump) the resulting solutionand the solubility of the components in the solution. Generally, theviscosity of the solution can determine the size of the particle in theresulting powder product.

Techniques and methods for spray-drying may be found in Perry's ChemicalEngineering Handbook, 6th Ed., R. H. Perry, D. W. Green & J. O. Maloney,eds., McGraw-Hill Book Co. (1984); and Marshall “Atomization andSpray-Drying” 50, Chem. Eng. Prog. Monogr. Series 2 (1954). In general,the spray-drying is conducted with an inlet temperature of from about40° C. to about 200° C., for example, from about 70° C. to about 150°C., preferably from about 40° C. to about 60° C., about 50° C. to about55° C., or about 80° C. to about 110° C., e.g., about 90° C. Thespray-drying is generally conducted with an outlet temperature of fromabout 20° C. to about 100° C., for example from about 25° C. to about30° C. (e.g., about 26° C.), about 40° C. to about 50° C., about 50° C.to about 65° C., e.g., about 56° C. to about 58° C.

Removal of the solvent or solvent mixture may require a subsequentdrying step, such as tray drying, fluid bed drying (e.g., from aboutroom temperature to about 100° C.), vacuum drying, microwave drying,rotary drum drying or biconical vacuum drying (e.g., from about roomtemperature to about 200° C.).

In one embodiment, the spray-drying is fluidized spray drying (FSD). Thesteps in FSD can include, for example: preparing a liquid feed solution(e.g., containing COMPOUND 2, and optionally a polymer(s) and/orsurfactant(s), dissolved or suspended in solvent(s)); atomizing (e.g.,with a pressure nozzle, a rotary atomizer or disk, two-fluid nozzle orother atomizing methods) the feed solution upon delivery into the dryingchamber of a spray dryer, e.g., operating in FSD mode; drying the feedsolution in the drying chamber with heated air or a heated gas (e.g.,nitrogen) to obtain a product, wherein larger particles of productseparate out, e.g., drop out, while fines are carried by a stream of airor gas up to the top of the drying chamber (e.g., by natural convection)and to a cyclone, and re-introducing (e.g., at the top of the dryingchamber or axially to the middle of the chamber) the fines into thedrying chamber, wherein the re-introduced fines can agglomerate withnewly formed product to generate an agglomerated product, wherein if theagglomerated product is large enough, it will separate out, if it is notlarge enough to separate out, the agglomerated product will be carriedby convection to the top of the chamber and to the cyclone andre-introduced into the chamber. This process repeats until anagglomerated product that is large enough to drop out is formed. Thefines can be re-introduced from the cyclone to the drying chamber via afeed pipe.

In some embodiments, rather than drying the feed solution with heatedair or a heated gas, the feed solution can instead be spray congealed,e.g., the chamber is at room temperature (e.g., 21±4° C.) or is cooled,e.g., cooled gas (e.g., nitrogen) is used for the process.

FSD can further include collecting the agglomerated product in a firstfluidizing chamber; which can be followed by discharging theagglomerated product from the first fluidizing chamber to a secondfluidizing chamber, wherein a post-drying process can occur.

The agglomerated product (e.g., that separates out in the dryingchamber) can then be transferred from the second fluidizing chamber to athird fluidizing chamber, where the agglomerated product is cooled. Theagglomerated product (e.g., a solid dispersion of an amorphous compound)can then be further processed. For example, the product can be directlycompressed. The product can optionally be blended with a surfactant,excipient, or pharmaceutically acceptable carrier, e.g., prior to directcompression. The product can optionally be further processed, e.g.,milled, granulated, blended, and/or mixed with a melt granulate,surfactant, excipient, and/or pharmaceutically acceptable carrier.

FSD can be performed in a commercial spray dryer operating in fluidizedspray dryer mode (FSD mode). FSD can be accomplished in either opencycle mode or closed cycle mode (e.g., the drying gas, e.g., nitrogen,is recycled). Examples of suitable spray dryers for use in FSD includedryers from Niro (e.g., the PSD line of spray driers manufactured byNiro: PHARMASD™; Chemical or SD line dryers). FSD can essentially beperformed in any spray dryer that is configured to allow for there-introduction of fines into the drying chamber.

Additional post drying, e.g., in a vacuum or fluidized bed dryer or adouble cone or biconical post-dryer or a tumble dryer, can be performedif needed/applicable to remove further solvents. In some embodiments, apost-drying step is performed.

To remove the solvent or solvent mixture, vacuum drying, spray drying,fluidized spray drying, tray drying, lyophilization, rotovapping, andother drying procedures may be applied. Applying any of these methodsusing appropriate processing parameters, according to this disclosure,would provide COMPOUND 2 in an amorphous state in the final soliddispersion product. Upon use of appropriate conditions (e.g., low outlettemperatures in the spray dryer, use of low boiling point solvents, useof heated gas) that result in a dispersion, e.g., powder, with desirableproperties (e.g., median particle size (d50) of 40-200 microns 9 e.g.,40-150 microns), powder bulk density of >0.2 g/ml (e.g., 0.2 to 0.5g/ml), or >0.25 g/ml, improved powder flowability (e.g., low cohesionforces, low interparticle internal friction); and/or dry powder with lowOVIs (Organic Volatile Impurities), e.g., below ICH limits and/or userspecifications), the dispersion can be directly compressed into a dosageform.

In some embodiments, the inlet temperature is between about 50° C. andabout 200° C., e.g., between about 60° C. and about 150° C., betweenabout 70° C. and about 100° C., between about 60° C. and about 95° C.,between about 65° C. and about 85° C., between about 70° C. and about90° C., between about 85° C. and about 95° C., or between about 70° C.and about 85° C.

In some embodiments, the outlet temperature is between about roomtemperature (e.g., USP room temperature (e.g., 21±4° C.)) and about 80°C., e.g., between about 25° C. and about 75° C., between about 30° C.and about 65° C., between about 35° C. and about 70° C., between about40° C. and about 65° C., between about 45° C. and about 60° C., betweenabout 35° C. and about 45° C., between about 35° C. and about 40° C., orbetween about 37° C. and about 40° C.

In some embodiments, the temperature set points of the fluidized beds(the temperature for each bed being selected independently from thetemperature selected for another bed) is between about room temperature(e.g., USP room temperature (e.g., 21±4° C.)) and about 100° C., e.g.,between about 30° C. and about 95° C., between about 40° C. and about90° C., between about 50° C. and about 80° C., between about 60° C. andabout 85° C., between about 65° C. and about 95° C., or between about80° C. and about 95° C.

FSD can be performed on a mixture containing COMPOUND 2. For example,FSD can be performed on a mixture containing COMPOUND 2, and one or morepolymer(s), and optionally one or more surfactant(s), and optionally oneor more additional excipients(s)) to obtain a solid dispersion ofamorphous COMPOUND 2 thereof, e.g., that can be directly compressed intoan oral dosage form (e.g., tablet). Alternatively, the dispersion can beblended with one or more excipients prior to compression.

In one embodiment, the process for preparing a solid dispersion ofCOMPOUND 2 comprises:

a) forming a mixture of COMPOUND 2, one or more polymer(s), and one ormore solvent(s); and

b) rapidly removing the solvent(s) from the solution to form a solidamorphous dispersion comprising COMPOUND 2 and the one or morepolymer(s). The one or more polymer(s) and one or more solvent(s) may beany of those disclosed herein.

In some embodiments, the solvent is removed by spray drying. In someembodiments the solid dispersion is tray dried using a convection traydryer. In some embodiments, the solid dispersion is screened.

In one embodiment, COMPOUND 2 is crystalline. In another embodiment,COMPOUND 2 is amorphous.

As would be appreciated by one of skill in the art, spray drying may bedone and is often done in the presence of an inert gas such as nitrogen.In certain embodiments, processes that involve spray drying may be donein the presence of a supercritical fluid involving carbon dioxide or amixture including carbon dioxide.

In another embodiment, the process for preparing a solid dispersion ofCOMPOUND 2 comprises:

a) forming a mixture of COMPOUND 2, a polymer, and a solvent; and

b) spray-drying the mixture to form a solid dispersion comprisingCOMPOUND 2 and the polymer.

Post-drying and/or polishing the wet spray dried dispersion to below ICHor given specifications for residual solvents can optionally beperformed.

These processes may be used to prepare the pharmaceutical compositionsdisclosed herein. The amounts and the features of the components used inthe processes may be as disclosed herein.

In some embodiments, the solvent comprises one or more volatilesolvent(s) to dissolve or suspend COMPOUND 2 and the polymer(s). In someembodiments, the one or more solvent(s) completely dissolves COMPOUND 2and the polymer(s).

In some embodiments, the one or more solvent(s) is a volatile solvent(e.g., methylene chloride, acetone, methanol, ethanol, chloroform,tetrahydrofuran (THF), or a mixture thereof). Examples of suitablevolatile solvents include those that dissolve or suspend thetherapeutically active compound either alone or in combination withanother co-solvent. In some embodiments, the solvent(s) completelydissolves the therapeutically active compound. In some embodiments, thesolvent is acetone. In some embodiments, the solvent is methanol.

In some embodiments, the solvent is a non-volatile solvent (e.g.,organic acids such as glacial acetic acid, dimethyl sulfoxide (DMSO),dimethylformamide (DMF), or water). In some embodiments, a non-volatilesolvent is a component in a solvent system. For example the non-volatilesolvent is present as a component in a solvent from about 1% to about20% w/w (e.g., from about 3% w/w to about 15% w/w, from about 4% w/w toabout 12% w/w, or from about 5% w/w to about 10% w/w).

In some embodiments, the solvent is a mixture of solvents. For example,the solvent can include from about 0% to about 30% acetone and fromabout 70% to about 100% methanol, or the solvent can include from about0% to about 40% acetone and from about 60% to about 100% methanol. Otherexemplary ratios of methanol to acetone include 80:20, 75:25, 70:30,60:40, 55:45, and 50:50.

In some embodiments, the solvent is a combination of solvents includingat least one non-volatile solvent. For example, the solvent is acombination of components that includes both a volatile solvent and anon-volatile solvent. In some embodiments, the solvent system is acombination of a volatile solvent or combination of solvents such asmethanol and acetone with a non-volatile solvent such as glacial aceticacid. For example, the solvent system comprises from about 40% to about80% methanol, from about 20% to about 35% acetone, and from about 1% toabout 15% glacial acetic acid (e.g., from about 50% to about 70%methanol, from about 25% to about 30% acetone, and from about 3% toabout 12% glacial acetic acid).

In some embodiments, the solvent system is a combination of a volatilesolvent or combination of solvents such as methanol and acetone with anon-volatile solvent such as water. For example, the solvent systemcomprises from about 40% to about 80% methanol, from about 20% to about35% acetone, and from about 0.1% to about 15% water (e.g., from about50% to about 70% methanol, from about 25% to about 30% acetone, and fromabout 1% to about 5% water).

In certain embodiments, the pharmaceutical compositions of the soliddispersion may be made by a process described herein. For example, asolid dispersion of: (a) COMPOUND 2 and (b) one or more polymer(s), andoptionally one or more surfactant(s) and optionally one or moreadditional excipient(s).

Pharmaceutical Compositions Containing Solid Dispersions of COMPOUND 2

In certain embodiments, provided herein are pharmaceutical compositions,comprising: (a) a solid dispersion, comprising COMPOUND 2 and a polymer;and (b) one or more pharmaceutically acceptable carrier(s). Examples ofpharmaceutically acceptable carriers are fillers, disintegrants, wettingagents, glidants, and lubricants.

In some embodiments, the pharmaceutical compositions may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, emulsions and aqueous suspensions,dispersions and solutions.

In some embodiments the pharmaceutical composition is a tablet.

In some embodiments the pharmaceutical composition comprises a directlycompressed dosage form of COMPOUND 2.

In some embodiments, the pharmaceutical composition also includes afiller. The filler can be, for example, microcrystalline cellulose,lactose, mannitol, ethyl cellulose, sorbitol, starch, sucrose, calciumphosphate, powdered cellulose, silicified microcrystalline cellulose,isomalt, or mixtures thereof. In some embodiments, the filler ismicrocrystalline cellulose.

In some embodiments, the filler is present in the pharmaceuticalcomposition in an amount of between about 10% w/w and 50% w/w (e.g.,between about 15% w/w and about 45% w/w; between about 20% w/w and about40% w/w; between about 25% w/w and about 35% w/w; or between about 28%w/w and about 32% w/w). In some embodiments, the filler is present inthe pharmaceutical composition in an amount of from about 20% w/w toabout 35% w/w, for example from about 25% w/w to about 34% w/w, or fromabout 26% w/w to about 33% w/w, or from about 27% w/w to about 32% w/w,for example, about 28% w/w, about 28.5% w/w, about 29% w/w, about 29.5%w/w about 30% w/w, about 30.5% w/w, about 31% w/w, or about 31.5% w/w.In some embodiments, the filler is present in the pharmaceuticalcomposition in an amount of about 29% w/w, about 29.1% w/w, about 29.2%w/w, about 29.3% w/w, about 29.4% w/w, about 29.5% w/w, about 29.6% w/w,about 29.7% w/w, about 29.8% w/w, about 29.9% w/w, or about 30% w/w. Insome embodiments, the filler is present in the pharmaceuticalcomposition in an amount of between about 25% w/w and about 35% w/w. Insome embodiments, the filler is present in the pharmaceuticalcomposition in an amount of about 29.5% w/w.

In some embodiments, the pharmaceutical composition also includes adisintegrant. The disintegrant can be, for example, colloidal silicondioxide, powdered cellulose, calcium silicate, crospovidone, calciumalginate, methyl cellulose, chitosan, carboxy methyl cellulose,croscarmellose sodium, carboxymethyl starch, sodium alginate, sodiumstarch glycolate, pregelatinized starch, or mixtures thereof. In someembodiments, the disintegrant is croscarmellose sodium.

In some embodiments, the disintegrant is present in the pharmaceuticalcomposition in an amount of between about 1% w/w and 15% w/w (e.g.,between about 3% w/w and about 12% w/w; between about 4% w/w and about10% w/w; between about 5% w/w and about 7% w/w; or between about 6% w/wand about 7% w/w). In some embodiments, the disintegrant is present inthe pharmaceutical composition in an amount of about 3% w/w, about 3.5%w/w, about 4% w/w, about 49.5% w/w about 5% w/w, about 5.5% w/w, about6% w/w, or about 6.5% w/w, about 7% w/w, about 7.5% w/w, about 8% w/w,about 8.5% w/w, about 9% w/w, about 9.5% w/w, or about 10% w/w. In someembodiments, the disintegrant is present in the pharmaceuticalcomposition in an amount of between about 5% w/w and about 7% w/w. Insome embodiments, the disintegrant is present in the pharmaceuticalcomposition in an amount of about 6% w/w.

In some embodiments, the pharmaceutical composition also includes awetting agent. The wetting agent can be, for example, sodium laurylsulfate, sodium dodecyl sulfate, polysorbates (such as Tween 20 andTween 80), poloxamers (such as Poloxamer 335 and Poloxamer 407),glyceryl monooleate, or mixtures thereof. In some embodiments, thewetting agent is sodium lauryl sulfate.

In some embodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of between about 0.1% w/w and 2% w/w (e.g.,between about 0.5% w/w and about 2% w/w; between about 0.5% w/w andabout 1.5% w/w; or between about 1% w/w and about 1.5% w/w). In someembodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of about 0.1% w/w, about 0.2% w/w, about 0.3%w/w, about 0.4% w/w about 0.5% w/w, about 0.6% w/w, about 0.7% w/w, orabout 0.8% w/w, about 0.9% w/w, about 1% w/w, about 1.1% w/w, about 1.2%w/w, about 1.3% w/w, about 1.4% w/w, about 1.5% w/w, about 1.6% w/w,about 1.7% w/w, about 1.8% w/w, about 1.9% w/w, or about 2% w/w. In someembodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of between about 0.5% w/w and about 1.5% w/w.In some embodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of about 1% w/w.

In some embodiments, the pharmaceutical composition also includes aglidant. The glidant can be, for example, silicon dioxide, colloidalsilicon dioxide, tribasic calcium phosphate, magnesium stearate,magnesium trisilicate, powdered cellulose, talc, starch, and mixturesthereof. In some embodiments, the glidant is colloidal silicon dioxide.

In some embodiments, the glidant is present in the pharmaceuticalcomposition in an amount of between about 0.1% w/w and 5% w/w (e.g.,between about 1% w/w and about 4% w/w; between about 1% w/w and about 3%w/w; or between about 1.5% w/w and about 2.5% w/w). In some embodiments,the glidant is present in the pharmaceutical composition in an amount ofabout 0.5% w/w, about 1% w/w, about 1.5% w/w, about 2% w/w about 2.5%w/w, about 3% w/w, about 3.5% w/w, or about 4% w/w, about 4.5% w/w, orabout 5% w/w. In some embodiments, the glidant is present in thepharmaceutical composition in an amount of about 1.1% w/w, about 1.2%w/w, about 1.3% w/w, about 1.4% w/w, about 1.5% w/w, about 1.6% w/w,about 1.7% w/w, about 1.8% w/w, about 1.9% w/w, about 2% w/w, 2.1% w/w,about 2.2% w/w, about 2.3% w/w, about 2.4% w/w, about 2.5% w/w, about2.6% w/w, about 2.7% w/w, about 2.8% w/w, about 2.9% w/w, or about 3%w/w. In some embodiments, the glidant is present in the pharmaceuticalcomposition in an amount of between about 1% w/w and about 3% w/w. Insome embodiments, the glidant is present in the pharmaceuticalcomposition in an amount of about 2% w/w.

In some embodiments, the pharmaceutical composition also includes alubricant. The lubricant can be, for example, magnesium stearate, talc,sodium stearyl fumarate, glyceryl behenate, hydrogenated vegetable oil,zinc stearate, calcium stearate, sucrose stearate, polyvinyl alcohol,magnesium lauryl sulfate, or mixtures thereof. In some embodiments, thelubricant is magnesium stearate.

In some embodiments, the lubricant is present in the pharmaceuticalcomposition in an amount of between about 0.1% w/w and 5% w/w (e.g.,between about 1% w/w and about 4% w/w; between about 1% w/w and about 3%w/w; or between about 1% w/w and about 2% w/w). In some embodiments, thelubricant is present in the pharmaceutical composition in an amount ofabout 0.5% w/w, about 1% w/w, about 1.5% w/w, about 2% w/w about 2.5%w/w, about 3% w/w, about 3.5% w/w, or about 4% w/w, about 4.5% w/w, orabout 5% w/w. In some embodiments, the lubricant is present in thepharmaceutical composition in an amount of about 0.1% w/w, about 0.2%w/w, about 0.3% w/w, about 0.4% w/w, about 0.5% w/w, about 0.6% w/w,about 0.7% w/w, about 0.8% w/w, about 0.9% w/w, about 1% w/w, about 1.1%w/w, about 1.2% w/w, about 1.3% w/w, about 1.4% w/w, about 1.5% w/w,about 1.6% w/w, about 1.7% w/w, about 1.8% w/w, about 1.9% w/w, about 2%w/w, 2.1% w/w, about 2.2% w/w, about 2.3% w/w, about 2.4% w/w, or about2.5% w/w. In some embodiments, the lubricant is present in thepharmaceutical composition in an amount of between about 0.5% w/w andabout 2.5% w/w. In some embodiments, the lubricant is present in thepharmaceutical composition in an amount of about 1.5% w/w.

In some embodiments, the solid dispersion makes up about 25% to 85% byweight of the total weight of the pharmaceutical composition. In someembodiments, the solid dispersion makes up about 50% to about 70% byweight of the total weight of the pharmaceutical composition.

In some embodiments, the COMPOUND 2 makes up about 15% to 45% of thetotal weight of the pharmaceutical composition, and the one or morepolymer(s) makes up about 15% to 45% of the total weight of thepharmaceutical composition.

In some embodiments, the COMPOUND 2 makes up about 20% w/w of thepharmaceutical composition, the one or more polymer(s) makes up about40% w/w of the pharmaceutical composition.

In some embodiments, the COMPOUND 2 makes up about 25% w/w of thepharmaceutical composition, the one or more polymer(s) makes up about35% w/w of the pharmaceutical composition.

In some embodiments, the COMPOUND 2 makes up about 30% w/w of thepharmaceutical composition, the one or more polymer(s) makes up about30% w/w of the pharmaceutical composition.

In some embodiments, the COMPOUND 2 makes up about 35% w/w of thepharmaceutical composition, the one or more polymer(s) makes up about25% w/w of the pharmaceutical composition.

In some embodiments, the solid dispersion makes up from between about50% w/w to about 70% w/w of the pharmaceutical composition, the fillermakes up from between about 25% w/w to about 35% w/w of thepharmaceutical composition, the disintegrant makes up from between about5% w/w to about 7% w/w of the pharmaceutical composition, the wettingagent makes up from between about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, the glidant makes up from between about 1%w/w to about 3% w/w of the pharmaceutical composition, the lubricantmakes up from between about 0.5% w/w to about 2.5% w/w of thepharmaceutical composition thereby totaling 100% by weight of thecomposition.

In some embodiments, the solid dispersion makes up about 60% w/w of thepharmaceutical composition, the filler makes up about 29.5% w/w of thepharmaceutical composition, the disintegrant makes up about 6% w/w ofthe pharmaceutical composition, the wetting agent makes up about 1% w/wof the pharmaceutical composition, the glidant makes up about 2% w/w ofthe pharmaceutical composition, the lubricant makes up about 1.5% w/w ofthe pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprises, frombetween about 25% w/w to about 35% w/w of COMPOUND 2 from between about25% w/w to about 35% w/w of hypromellose acetate succinate (HPMCAS),from between about 25% w/w to about 35% w/w of microcrystallinecellulose, from between about 5% w/w to about 7% w/w croscarmellosesodium, from between about 0.5% w/w to about 1.5% w/w sodium laurylsulfate, about from between about 1% w/w to about 3% w/w colloidalsilicon dioxide, and from between about 0.5% w/w to about 2.5% w/w ofmagnesium stearate, thereby totaling 100% by weight of the composition.

In some embodiments, the pharmaceutical composition comprises, about 30%w/w of COMPOUND 2 about 30% w/w of hypromellose acetate succinate(HPMCAS), about 29.5% w/w of microcrystalline cellulose, about 6% w/wcroscarmellose sodium, about 1% w/w sodium lauryl sulfate, about 2% w/wcolloidal silicon dioxide, and about 1.5% w/w of magnesium stearate.

In some embodiments, the solid dispersion, filler, disintegrant, wettingagent, glidant, and lubricant are added intragranularly. In someembodiments, an additional amount of the filler, disintegrant, glidant,and lubricant are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersion makesup from about 50% w/w to about 70% w/w of the pharmaceuticalcomposition, the filler makes up from about 18% w/w to about 26% w/w ofthe pharmaceutical composition, disintegrant makes up from about 2% w/wto about 6% w/w of the pharmaceutical composition, wetting agent makesup from about 0.5% w/w to about 1.5% w/w of the pharmaceuticalcomposition, glidant makes up from about 0.5% w/w to about 1.5% w/w ofthe pharmaceutical composition, and lubricant makes up from about 0.25%w/w to about 1% w/w of the pharmaceutical composition.

In some embodiments, a the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount of thefiller makes up from about 4% w/w to about 12% w/w of the pharmaceuticalcomposition, an additional amount of the disintegrant makes up fromabout 1% w/w to about 3% w/w of the pharmaceutical composition, anadditional amount of the glidant makes up from about 0.5% w/w to about1.5% w/w of the pharmaceutical composition, and an additional amount ofthe lubricant makes up from about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, and are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersion makesup about 60% w/w of the pharmaceutical composition, the filler makes upabout 21.5% w/w of the pharmaceutical composition, disintegrant makes upabout 4% w/w of the pharmaceutical composition, wetting agent makes upabout 1% w/w of the pharmaceutical composition, glidant makes up about1% w/w of the pharmaceutical composition, and lubricant makes up about0.5% w/w of the pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount of thefiller makes up about 8% w/w of the pharmaceutical composition, anadditional amount of the disintegrant makes up about 2% w/w of thepharmaceutical composition, an additional amount of the glidant makes upabout 1% w/w of the pharmaceutical composition, and an additional amountof the lubricant makes up about 1% w/w of the pharmaceuticalcomposition, and are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersioncomprising COMPOUND 2 and hypromellose acetate succinate (HPMCAS), makesup from about 50% w/w to about 70% w/w of the pharmaceuticalcomposition, microcrystalline cellulose makes up from about 18% w/w toabout 26% w/w of the pharmaceutical composition, croscarmellose sodiummakes up from about 2% w/w to about 6% w/w of the pharmaceuticalcomposition, sodium lauryl sulfate makes up from about 0.5% w/w to about1.5% w/w of the pharmaceutical composition, colloidal silicon dioxidemakes up from about 0.5% w/w to about 1.5% w/w of the pharmaceuticalcomposition, and magnesium stearate makes up from about 0.25% w/w toabout 1% w/w of the pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount ofmicrocrystalline cellulose makes up from about 4% w/w to about 12% w/wof the pharmaceutical composition, an additional amount ofcroscarmellose sodium makes up from about 1% w/w to about 3% w/w of thepharmaceutical composition, an additional amount of colloidal silicondioxide makes up from about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, and an additional amount of magnesiumstearate makes up from about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, and are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersioncomprising COMPOUND 2 and hypromellose acetate succinate (HPMCAS), makesup about 60% w/w of the pharmaceutical composition, microcrystallinecellulose makes up about 21.5% w/w of the pharmaceutical composition,croscarmellose sodium makes up about 4% w/w of the pharmaceuticalcomposition, sodium lauryl sulfate makes up about 1% w/w of thepharmaceutical composition, colloidal silicon dioxide makes up about 1%w/w of the pharmaceutical composition, and magnesium stearate makes upabout 0.5% w/w of the pharmaceutical composition.

In some embodiments, a the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount ofmicrocrystalline cellulose makes up about 8% w/w of the pharmaceuticalcomposition, an additional amount of croscarmellose sodium makes upabout 2% w/w of the pharmaceutical composition, an additional amount ofcolloidal silicon dioxide makes up about 1% w/w of the pharmaceuticalcomposition, and an additional amount of magnesium stearate makes upabout 1% w/w of the pharmaceutical composition, and are addedextragranularly.

Methods of Use

It has been observed that the mutation status of FLT3 is associated withresponses in cancer characterized by the presence of a mutant allele ofIDH2 when treated with COMPOUND 1 and in cancer characterized by thepresence of a mutant allele of IDH1 when treated with COMPOUND 2. Whilenot intending to be bound by any particular theory of operation, somaticmutations in FLT3, for example FLT-ITD mutation, may be associated withresistance to treatment with COMPOUND 1 in AML characterized by thepresence of a mutant allele of IDH2, and resistance to treatment withCOMPOUND 2 in AML characterized by the presence of a mutant allele ofIDH1.

In one embodiment, the methods provided herein encompass treating,preventing, or managing cancer in a subject, wherein the cancer ischaracterized by the presence of a mutant allele of IDH2 and the absenceof a FLT3 mutation by administering a therapeutically effective amountof an IDH2 inhibitor. In one embodiment, the IDH2 inhibitor is COMPOUND1.

In one embodiment, the methods provided herein encompass treating,preventing, or managing cancer in a subject, wherein the cancer ischaracterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation by administering a therapeutically effective amountof an IDH1 inhibitor. In one embodiment, the an IDH1 inhibitor isCOMPOUND 2.

In one embodiment, provided herein is a method of treating, preventing,or managing solid tumors in a subject, wherein the solid tumor ischaracterized by the presence of a mutant allele of IDH2 and the absenceof a FLT3 mutation by administering a therapeutically effective amountof COMPOUND 1. In one embodiment, the solid tumor is an advanced solidtumor.

In one embodiment, provided herein is a method of treating, preventing,or managing solid tumors in a subject, wherein the solid tumor ischaracterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation by administering a therapeutically effective amountof COMPOUND 2. In one embodiment, the solid tumor is an advanced solidtumor.

In one embodiment, provided herein is a method of treating, preventing,or managing hematological malignancies in a subject, wherein thehematological malignancy is characterized by the presence of a mutantallele of IDH2 and the absence of a FLT3 mutation by administering atherapeutically effective amount of COMPOUND 1. In one embodiment, thehematologic malignancy is an advanced hematologic malignancy. In oneembodiment, the hematologic malignancy is AML.

In one embodiment, provided herein is a method of treating, preventing,or managing hematological malignancies in a subject, wherein thehematological malignancy is characterized by the presence of a mutantallele of IDH1 and the absence of a FLT3 mutation by administering atherapeutically effective amount of COMPOUND 2. In one embodiment, thehematologic malignancy is an advanced hematologic malignancy. In oneembodiment, the hematologic malignancy is AML.

In certain embodiments, the methods encompass treating, preventing, ormanaging cancer in a subject, wherein the cancer is characterized by thepresence of a mutant allele of IDH2 and a mutant FLT3 by administering atherapeutically effective amount of an IDH2 inhibitor, such as COMPOUND1, in combination with a therapeutically effective amount of one or morecompounds that target the FLT3 pathway. In one embodiment, the compoundthat targets the FLT3 pathway is a FLT3 inhibitor selected fromquizartinib (AC220), sunitinib (SU11248), sorafenib (BAY 43-9006),midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596),PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449,famitinib and DCC-2036.

In certain embodiments, the methods encompass treating, preventing, ormanaging cancer in a subject, wherein the cancer is characterized by thepresence of a mutant allele of IDH1 and a mutant FLT3 by administering atherapeutically effective amount of an IDH1 inhibitor, such as COMPOUND2, in combination with a therapeutically effective amount of one or morecompounds that target the FLT3 pathway. In one embodiment, the compoundthat targets the FLT3 pathway is a FLT3 inhibitor selected fromquizartinib (AC220), sunitinib (SU11248), sorafenib (BAY 43-9006),midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596),PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449,famitinib and DCC-2036.

In certain embodiments, the methods encompass treating, preventing, ormanaging a solid tumor in a subject, wherein the solid tumor ischaracterized by the presence of a mutant allele of IDH2 and a mutantFLT3 by administering a therapeutically effective amount of an IDH2inhibitor, such as COMPOUND 1 in combination with a therapeuticallyeffective amount of one or more compounds that the target FLT3 pathway.In one embodiment, the compound that targets the FLT3 pathway is a FLT3inhibitor selected from quizartinib (AC220), sunitinib (SU11248),sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib (CEP-701),crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534),ASP2215, KW-2449, famitinib and DCC-2036.

In certain embodiments, the methods encompass treating, preventing, ormanaging a solid tumor in a subject, wherein the solid tumor ischaracterized by the presence of a mutant allele of IDH1 and a mutantFLT3 by administering a therapeutically effective amount of an IDH1inhibitor, such as COMPOUND 2 in combination with a therapeuticallyeffective amount of one or more compounds that target the FLT3 pathway.In one embodiment, the compound that targets the FLT3 pathway is a FLT3inhibitor selected from quizartinib (AC220), sunitinib (SU11248),sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib (CEP-701),crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534),ASP2215, KW-2449, famitinib and DCC-2036.

In one embodiment, the solid tumor is an advanced solid tumor.

In certain embodiments, the methods encompass treating, preventing, ormanaging a hematologic malignancy in a subject, wherein thehematological malignancy is characterized by the presence of a mutantallele of IDH2 and a mutant FLT3 by administering a therapeuticallyeffective amount of an IDH2 inhibitor, such as COMPOUND 1 in combinationwith a therapeutically effective amount of one or more compounds thattarget the FLT3 pathway. In one embodiment, the compound that targetsthe FLT3 pathway is a FLT3 inhibitor selected from quizartinib (AC220),sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412),lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028,ponatinib (AP24534), ASP2215, KW-2449, famitinib and DCC-2036.

In certain embodiments, the methods encompass treating, preventing, ormanaging a hematologic malignancy in a subject, wherein thehematological malignancy is characterized by the presence of a mutantallele of IDH1 and a mutant FLT3 by administering a therapeuticallyeffective amount of an IDH1 inhibitor, such as COMPOUND 2 in combinationwith a therapeutically effective amount of one or more compounds thattarget the FLT3 pathway. In one embodiment, the compound that targetsthe FLT3 pathway is a FLT3 inhibitor selected from quizartinib (AC220),sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412),lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028,ponatinib (AP24534), ASP2215, KW-2449, famitinib and DCC-2036.

In one embodiment, the hematologic malignancy is an advanced hematologicmalignancy. In one embodiment, the hematologic malignancy is AML.

In certain embodiments, methods provided herein comprise contacting acancer cell in or from a subject, such as a patient, wherein the cancercell is characterized by the presence of a mutant allele of IDH2 and theabsence of a FLT3 mutation, with a therapeutically effective amount ofCOMPOUND 1. In certain embodiments, methods provided herein comprisecontacting a cancer cell in or from a subject, such as a patient,wherein the cancer cell is characterized by the presence of a mutantallele of IDH1 and the absence of a FLT3 mutation, with atherapeutically effective amount of COMPOUND 2. The contacting can be invitro, in vivo, or ex vivo. In one embodiment, the method comprisescontacting the cancer cell in vivo.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with an IDH1 inhibitor, comprising: (a)obtaining a biological sample from a subject having cancer; (b)screening the biological sample for an IDH1 mutation and a FLT3mutation, for example FLT3-ITD mutation; and (c) if the cancer ischaracterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation, identifying the subject as a cancer subject suitablefor treatment with an IDH1 inhibitor. In another embodiment, thesubjects identified as cancer subjects suitable for treatment with anIDH1 inhibitor are treated with an IDH1 inhibitor.

In one embodiment, provided is an IDH1 inhibitor for use in a method fortreating cancer in a cancer subject, wherein the cancer subject has beenidentified by the method of identifying the cancer subject suitable fortreatment with an IDH1 inhibitor, comprising: (a) obtaining a biologicalsample from a subject having cancer; (b) screening the biological samplefor an IDH1 mutation and a FLT3 mutation; and (c) if the cancer ischaracterized by the presence of a mutant allele of IDH1 and the absenceof a FLT3 mutation, identifying the subject as a cancer subject suitablefor treatment with an IDH1 inhibitor.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with an IDH2 inhibitor, comprising: (a)obtaining a biological sample from a subject having cancer; (b)screening the biological sample for an IDH2 mutation and a FLT3mutation, for example FLT3-ITD mutation; and (c) if the cancer ischaracterized by the presence of a mutant allele of IDH2 and the absenceof a FLT3 mutation, identifying the subject as a cancer subject suitablefor treatment with an IDH2 inhibitor. In another embodiment, thesubjects identified as cancer subjects suitable for treatment with anIDH2 inhibitor are treated with an IDH2 inhibitor.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with COMPOUND 1, comprising: (a)obtaining a biological sample from a subject having cancer; (b)screening the biological sample for an IDH2 mutation and a FLT3mutation, for example FLT3-ITD mutation; and (c) if the cancer ischaracterized by the presence of a mutant allele of IDH2 and the absenceof a FLT3 mutation, identifying the subject as a cancer subject suitablefor treatment with COMPOUND 1. In another embodiment, the subjectsidentified as cancer subjects suitable for treatment with COMPOUND 1 aretreated with COMPOUND 1.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with COMPOUND 2, comprising: (a)obtaining a biological sample from a subject having cancer; (b)screening the biological sample for an IDH1 mutation and a FLT3mutation; and (c) if the cancer is characterized by the presence of amutant allele of IDH1 the absence of a FLT3 mutation, identifying thesubject as a cancer subject suitable for treatment with COMPOUND 2. Inanother embodiment, the subjects identified as cancer subjects suitablefor treatment with COMPOUND 2 are treated with COMPOUND 2.

Provided is also COMPOUND 2 for use in a method for treating cancer in acancer subject, wherein the cancer subject has been identified by themethod of identifying a cancer subject suitable for treatment withCOMPOUND 2, comprising: (a) obtaining a biological sample from a subjecthaving cancer; (b) screening the biological sample for an IDH1 mutationand a FLT3 mutation; and (c) if the cancer is characterized by thepresence of a mutant allele of IDH1 and the absence of a FLT3 mutation,identifying the subject as a cancer subject suitable for treatment withCOMPOUND 2.

In another embodiment, provided herein is a method for identifying oneor more cancer subjects suitable for treatment with an IDH2 inhibitorfrom a plurality of cancer subjects with a cancer characterized by thepresence of a mutant allele of IDH2. The method comprises identifyingone or more cancer subjects with a cancer characterized by the absenceof a FLT3 mutation from the plurality of cancer subjects suitable fortreatment with an IDH2 inhibitor. In one embodiment, one or moresuitable subjects are treated with an IDH2 inhibitor.

In another embodiment, provided herein is a method for identifying oneor more cancer subjects suitable for treatment with an IDH1 inhibitorfrom a plurality of cancer subjects, with a cancer characterized by thepresence of a mutant allele of IDH1. The method comprises identifyingone or more cancer subjects with a cancer characterized by the absenceof a FLT3 mutation from the plurality of cancer subjects suitable fortreatment with an IDH1 inhibitor. In one embodiment, one or moresuitable subjects are treated with an IDH1 inhibitor.

Provided is also the IDH1 inhibitor for use in a method for treatingcancer in one or more cancer subjects with a cancer characterized by thepresence of a mutant allele of IDH1, wherein the one or more cancersubjects are identified by the method comprising identifying one or morecancer subjects with a cancer characterized by the absence of a FLT3mutation from the plurality of cancer subjects suitable for treatmentwith an IDH1 inhibitor.

In another embodiment, provided herein is a method for identifying oneor more cancer subjects suitable for treatment with COMPOUND 1 from aplurality of cancer subjects, with a cancer characterized by thepresence of a mutant allele of IDH2. The method comprises identifyingone or more cancer subjects with a cancer characterized by the absenceof a FLT3 mutation from the plurality of cancer subjects suitable fortreatment with COMPOUND 1. In one embodiment, one or more suitablesubjects are treated with COMPOUND 1.

In another embodiment, provided herein is a method for identifying oneor more cancer subjects suitable for treatment with COMPOUND 2 from aplurality of cancer subjects with a cancer characterized by the presenceof a mutant allele of IDH1. The method comprises identifying one or morecancer subjects characterized by the absence of a FLT3 mutation from theplurality of cancer subjects suitable for treatment with COMPOUND 2. Inone embodiment, one or more suitable subjects are treated with COMPOUND2.

Provided is also an IDH1 inhibitor for use in a method for treatingcancer in one or more cancer subjects with a cancer characterized by thepresence of a mutant allele of IDH1. In an embodiment, the one or morecancer subjects are identified by the method comprising identifying oneor more cancer subjects with a cancer characterized by the absence of aFLT3 mutation from the plurality of cancer subjects suitable fortreatment with an IDH1 inhibitor. In one embodiment, one or moresuitable subjects are treated with COMPOUND 2.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with a combination of an IDH1 inhibitorand a FLT3 pathway inhibitor, comprising: (a) obtaining a biologicalsample from a subject having cancer; (b) screening the biological samplefor an IDH1 mutation and a FLT3 mutation, for example FLT3-ITD mutation;and (c) if the cancer is characterized by the presence of a mutantallele of IDH1 and a mutant FLT3, for example FLT3-ITD, identifying thesubject as a cancer subject suitable for treatment with a combinationtherapy with an IDH1 inhibitor and a FLT3 pathway inhibitor. In anotherembodiment, the subjects identified as cancer subjects suitable fortreatment with the combination therapy are treated with a combination ofan IDH1 inhibitor and a FLT3 pathway inhibitor. In one embodiment, theFLT3 pathway inhibitor is selected from quizartinib (AC220), sunitinib(SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib(CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib(AP24534), ASP2215, KW-2449, famitinib and DCC-2036.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with a combination of an IDH2 inhibitorand a FLT3 pathway inhibitor, comprising: (a) obtaining a biologicalsample from a subject having cancer; (b) screening the biological samplefor an IDH2 mutation and a FLT3 mutation, for example FLT3-ITD mutation;and (c) if the cancer is characterized by the presence of a mutantallele of IDH2 and a mutant FLT3, for example FLT3-ITD, identifying thesubject as a cancer subject suitable for treatment with a combinationtherapy with an IDH2 inhibitor and a FLT3 pathway inhibitor. In anotherembodiment, the subjects identified as cancer subjects suitable fortreatment with the combination therapy are treated with a combination ofan IDH2 inhibitor and a FLT3 pathway inhibitor.

In one embodiment, the FLT3 inhibitor is selected from quizartinib(AC220), sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin(PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397,E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449, famitinib andDCC-2036.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with a combination of COMPOUND 1 and aFLT3 pathway inhibitor, comprising: (a) obtaining a biological samplefrom a subject having cancer; (b) screening the biological sample for anIDH2 mutation and a FLT3 mutation, for example FLT3-ITD mutation; and(c) if the cancer is characterized by the presence of a mutant allele ofIDH2 and a mutant FLT3, for example FLT3-ITD, identifying the subject asa cancer subject suitable for treatment with a combination therapy withCOMPOUND 1 and a FLT3 pathway inhibitor. In another embodiment, thesubjects identified as cancer subjects suitable for treatment with thecombination therapy are treated with a combination of COMPOUND 1 and aFLT3 pathway inhibitor. In one embodiment, the FLT3 inhibitor isselected from quizartinib (AC220), sunitinib (SU11248), sorafenib (BAY43-9006), midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib(CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215,KW-2449, famitinib and DCC-2036.

In one embodiment, provided herein is a method of identifying a cancersubject suitable for treatment with a combination of COMPOUND 2 and aFLT3 pathway inhibitor, comprising: (a) obtaining a biological samplefrom a subject having cancer; (b) screening the biological sample for anIDH1 mutation and a FLT3 mutation; and (c) if the cancer ischaracterized by the presence of a mutant allele of IDH1 and a mutantFLT3, identifying the subject as a cancer subject suitable for treatmentwith a combination therapy with COMPOUND 2 and a FLT3 pathway inhibitor.In another embodiment, the subjects identified as cancer subjectssuitable for treatment with the combination therapy are treated with acombination of COMPOUND 2 and a FLT3 pathway inhibitor.

Provided is also a combination of an IDH1 inhibitor and a FLT3 pathwayinhibitor for use in a method for treating cancer in a cancer subject,wherein the cancer subject has been identified by the method ofidentifying a cancer subject suitable for treatment with a combinationof an IDH1 inhibitor and a FLT3 pathway inhibitor, comprising: (a)obtaining a biological sample from a subject having cancer; (b)screening the biological sample for an IDH1 mutation and a FLT3mutation; and (c) if the cancer is characterized by the presence of amutant allele of IDH1 and a mutant FLT3, identifying the subject as acancer subject suitable for treatment with a combination therapy with anIDH1 inhibitor and a FLT3 pathway inhibitor. In an embodiment, the IDH1inhibitor is COMPOUND2.

In one embodiment, the FLT3 inhibitor is selected from quizartinib(AC220), sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin(PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397,E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449, famitinib andDCC-2036.

In another embodiment, provided herein is a method for identifying oneor more cancer subjects suitable for treatment with a combinationtherapy with an IDH2 inhibitor, for example COMPOUND 1, and FLT3 pathwayinhibitor, for example, quizartinib (AC220), sunitinib (SU11248),sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib (CEP-701),crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534),ASP2215, KW-2449, famitinib or DCC-2036, from a plurality of cancersubjects. The method comprises identifying one or more cancer subjects,wherein the cancer is characterized by the presence of a mutant alleleof IDH2 and a mutant FLT3 from the plurality of cancer subjects suitablefor treatment with a combination therapy with an IDH2 inhibitor, forexample COMPOUND 1, and FLT3 pathway inhibitor, for example, quizartinib(AC220), sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin(PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397,E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449, famitinib orDCC-2036. In one embodiment, one or more suitable subjects are treatedwith a combination of an IDH2 inhibitor, for example COMPOUND 1, andFLT3 pathway inhibitor, for example, quizartinib (AC220), sunitinib(SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib(CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib(AP24534), ASP2215, KW-2449, famitinib or DCC-2036.

In another embodiment, provided herein is a method for identifying oneor more cancer subjects suitable for treatment with a combinationtherapy with an IDH1 inhibitor, for example, COMPOUND 2, and FLT3pathway inhibitor, for example, FLT3 inhibitor selected from quizartinib(AC220), sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin(PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397,E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449, famitinib andDCC-2036, from a plurality of cancer subjects. The method comprisesidentifying one or more cancer subjects, wherein the cancer ischaracterized by the presence of a mutant allele of IDH1 and a mutantFLT3 from the plurality of cancer subjects suitable for treatment with acombination therapy with an IDH1 inhibitor, for example COMPOUND 2, andFLT3 pathway inhibitor, for example, FLT3 inhibitor selected fromquizartinib (AC220), sunitinib (SU11248), sorafenib (BAY 43-9006),midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596),PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449,famitinib and DCC-2036. In one embodiment, one or more suitable subjectsare treated with a combination of an IDH1 inhibitor, for exampleCOMPOUND 2, and FLT3 pathway inhibitor, for example, FLT3 inhibitorselected from quizartinib (AC220), sunitinib (SU11248), sorafenib (BAY43-9006), midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib(CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215,KW-2449, famitinib and DCC-2036.

Provided is also a combination of an IDH1 inhibitor, for exampleCOMPOUND 2, and FLT3 pathway inhibitor, for example, quizartinib(AC220), sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin(PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397,E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449, famitinib orDCC-2036 for use in a method for treating cancer in one or more cancersubjects, wherein the one or more cancer subjects are identified by themethod comprising identifying one or more cancer subjects, wherein thecancer is characterized by the presence of a mutant allele of IDH1 and amutant FLT3 from the plurality of cancer subjects suitable for treatmentwith a combination therapy with an IDH1 inhibitor, for example COMPOUND2, and a FLT3 pathway inhibitor, for example, quizartinib (AC220),sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412),lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028,ponatinib (AP24534), ASP2215, KW-2449, famitinib or DCC-2036.

In one embodiment, one or more cancer subjects with a cancercharacterized by the presence of a FLT3 mutation are not treated with anIDH1 inhibitor, for example COMPOUND 2. In one embodiment, one or morecancer subjects with a cancer characterized by the presence of a mutantFLT3 are not treated with an IDH2 inhibitor, for example COMPOUND 1.

In certain embodiments, the inhibitory activity of COMPOUND 2 againstIDH1 mutants (e.g., IDH1 R132H, IDH1 R132C, IDH1 R132L, IDH1 R132V, IDH1R132S or IDH1 R132G) can be tested by methods described in Example A ofPCT Publication No. WO 2013/107291 and US Publication No. US2013/0190249. In certain embodiment, the inhibitory activity of COMPOUND1 against IDH2 mutants can be tested by methods described in USPublication No. US2013/0190287, hereby incorporated by reference intheir entireties, or analogous methods.

In some embodiments of the methods described herein, the hematologicmalignancy is acute myelogenous leukemia (AML), myelodysplastic syndrome(MDS), chronic myelomonocytic leukemia (CMML), myeloid sarcoma, multiplemyeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, each characterized by the presence of a mutantallele of IDH2. In other embodiments of the methods described herein thehematologic malignancy is acute myelogenous leukemia (AML),myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), each characterized by thepresence of a mutant allele of IDH1. In one embodiment, the hematologicmalignancy is an advanced hematologic malignancy.

In some embodiments of the methods described herein, the hematologicmalignancy is acute myelogenous leukemia (AML) characterized by thepresence of a mutant allele of IDH1. In some embodiments of the methodsdescribed herein, the acute myelogenous leukemia (AML) is relapse orrefractory AML, characterized by the presence of a mutant allele ofIDH1. In some embodiments of the methods described herein, thehematologic malignancy is myelodysplastic syndrome (MDS) characterizedby the presence of a mutant allele of IDH1. In some embodiments of themethods described herein, the hematologic malignancy is chronicmyelomonocytic leukemia (CMML) characterized by the presence of a mutantallele of IDH1. In some embodiments of the methods described herein, thehematologic malignancy is myeloid sarcoma characterized by the presenceof a mutant allele of IDH1. In some embodiments of the methods describedherein, the hematologic malignancy is lymphoma (e.g., T-cell lymphoma orB-cell lymphoma) characterized by the presence of a mutant allele ofIDH1. In some embodiments of the methods described herein, thehematologic malignancy is angioimmunoblastic T-cell lymphoma (AITL)characterized by the presence of a mutant allele of IDH1. In someembodiments of the methods described herein, the hematologic malignancyis blastic plasmacytoid dendritic cell neoplasm characterized by thepresence of a mutant allele of IDH1.

In one embodiment of the methods provided herein the solid tumor isglioma, melanoma, chondrosarcoma, cholangiocarcinoma (e.g., glioma),angioimmunoblastic T-cell lymphoma (AITL), sarcoma, or non small celllung cancer, each characterized by the presence of a mutant allele ofIDH2.

In one embodiment of the methods provided herein the solid tumor isglioma, melanoma, chondrosarcoma, cholangiocarcinoma (includingintrahepatic cholangiocarcinoma (IHCC), prostate cancer, colon cancer,or non-small cell lung cancer (NSCLC), each characterized by thepresence of a mutant allele of IDH1.

In one embodiment of the methods provided herein the solid tumor isglioma, characterized by the presence of a mutant allele of IDH1. In oneembodiment of the methods provided herein the solid tumor is melanomacharacterized by the presence of a mutant allele of IDH1. In oneembodiment of the methods provided herein the solid tumor chondrosarcomacharacterized by the presence of a mutant allele of IDH1. In oneembodiment of the methods provided herein the solid tumor ischolangiocarcinoma (e.g., glioma) characterized by the presence of amutant allele of IDH1. In one embodiment of the methods provided hereinthe solid tumor is angioimmunoblastic T-cell lymphoma (AITL)characterized by the presence of a mutant allele of IDH1. In oneembodiment of the methods provided herein the solid tumor is sarcomacharacterized by the presence of a mutant allele of IDH1. In oneembodiment of the methods provided herein the solid tumor is non smallcell lung cancer characterized by the presence of a mutant allele ofIDH1.

In one embodiment, the malignancy to be treated is characterized by amutant allele of IDH1 or IDH2, wherein the IDH1 or IDH2 mutation resultsin a new ability of the enzyme to catalyze the NAPH dependent reductionof a ketoglutarate to R(−)-2-hydroxyglutarate in a patient. In oneaspect of this embodiment, the mutant IDH1 has an R132X mutation. In oneaspect of this embodiment, the R132X mutation is selected from R132H,R132C, R132L, R132V, R132S and R132G. In another aspect, the R132Xmutation is R132H or R132C. In yet another aspect, the R132X mutation isR132H. In one aspect of this embodiment, the mutant IDH2 has an R140Xmutation. In another aspect of this embodiment, the R140X mutation is aR140Q mutation. In another aspect of this embodiment, the R140X mutationis a R140W mutation. In another aspect of this embodiment, the R140Xmutation is a R140L mutation. In another aspect of this embodiment, themutant IDH2 has an R172X mutation. In another aspect of this embodiment,the R172X mutation is a R172K mutation. In another aspect of thisembodiment, the R172X mutation is a R172G mutation.

In one embodiment, a malignancy can be analyzed by sequencing cellsamples to determine the presence and specific nature of (e.g., thechanged amino acid present at) a mutation at amino acid 132 of IDH1. Inone embodiment, a malignancy can be analyzed by sequencing cell samplesto determine the presence and specific nature of (e.g., the changedamino acid present at) a mutation at amino acid 140 and/or 172 of IDH2.

Without being bound by theory, applicants believe that mutant alleles ofIDH1 wherein the IDH1 mutation results in a new ability of the enzyme tocatalyze the NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate, and in particular R132H mutations of IDH1,characterize a subset of all types of cancers, without regard to theircellular nature or location in the body. Thus, COMPOUND 2, and methodsdescribed herein are useful to treat an hematologic malignancy,including an advanced hematologic malignancy, such as acute myelogenousleukemia (AML), myelodysplastic syndrome (MDS), myeloproliferativeneoplasms (MPN), chronic myelomonocytic leukemia (CMML), B-acutelymphoblastic leukemias (B-ALL), or lymphoma (e.g., T-cell lymphoma),each characterized by the presence of a mutant allele of IDH1 impartingsuch activity and in particular an IDH1 R132H or R132C mutation.

In another aspect, COMPOUND 2 and methods described herein are useful totreat a solid tumor, such as glioma, melanoma, chondrosarcoma,cholangiocarcinoma (including intrahepatic cholangiocarcinoma (IHCC),prostate cancer, colon cancer, or non-small cell lung cancer (NSCLC),each characterized by the presence of a mutant allele of IDH1 impartingsuch activity and in particular an IDH1 R132H or R132C mutation.

In one embodiment, the advanced hematologic malignancy to be treated isAML. In some embodiments, the AML is relapsed and/or refractory. Inother embodiments, the AML is untreated. In some embodiments, the AML isrelapsed and/or refractory in patients 60 years of age and older. Insome embodiments, the AML is untreated in patients 60 years of age andolder. In some embodiments, the AML is relapsed and/or refractory inpatients under 60 years of age. In one embodiment, COMPOUND 2 isadministered as a first line treatment for AML. In one embodiment,COMPOUND 2 is administered as a second line, third line, or fourth linetreatment for AML. In one embodiment, COMPOUND 2 is administered after afirst line treatment for AML. In one embodiment, COMPOUND 2 isadministered after a second line, third line, or fourth line treatmentfor AML. In one embodiment, COMPOUND 2 is administered after a firstrelapse. In one embodiment, COMPOUND 2 is administered after primaryinduction failure. In one embodiment, COMPOUND 2 is administered afterre-induction failure. In one embodiment, administration of COMPOUND 2can occur prior to, during, or after transplant. In one embodiment,COMPOUND 2 is administered after a relapse that is post-transplant. Inone embodiment, the AML presentation is subsequent to MPD. In oneembodiment, the AML presentation is subsequent to MDS and CMML.

In another aspect, without being bound by theory, applicants have foundthat mutant alleles of IDH2, wherein the IDH2 mutation results in a newability of the enzyme to catalyze the NAPH dependent reduction of aketoglutarate to R(−) 2 hydroxyglutarate, and in particular R140Q and/orR172K mutations of IDH2, characterize a subset of all types of cancers,without regard to their cellular nature or location in the body. Thus,the compounds, compositions and methods provided herein are useful totreat any type of cancer that is characterized by the presence of amutant allele of IDH2 imparting such activity and in particular an IDH2R140Q and/or R172K mutation.

Thus, COMPOUND 1, and methods described herein are useful to treat anhematologic malignancy, including an advanced hematologic malignancy,such as acute myelogenous leukemia (AML), myelodysplastic syndrome(MDS), chronic myelomonocytic leukemia (CMML), myeloid sarcoma, multiplemyeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, each characterized by the presence of a mutantallele of IDH2 imparting such activity and in particular an IDH2 R140Qand/or R172K mutation.

In another aspect, COMPOUND 2 and methods described herein are useful totreat a solid tumor, such as glioma, melanoma, chondrosarcoma,cholangiocarcinoma (e.g., glioma), angioimmunoblastic T-cell lymphoma(AITL), sarcoma, or non small cell lung cancer, each characterized bythe presence of a mutant allele of IDH 2 imparting such activity and inparticular IDH2 R140Q and/or R172K mutation.

In one embodiment the malignancy is a tumor wherein at least 30, 40, 50,60, 70, 80 or 90% of the tumor cells carry an IDH1 mutation, and inparticular an an IDH1 R132H or R132C mutation, or an IDH2 mutation, andin particular an IDH2 R140Q, R140W, or R140L and/or R172K or R172Gmutation, at the time of diagnosis or treatment.

In one embodiment, the efficacy of treatment of malignancy is monitoredby measuring the levels of 2HG in the subject. Typically levels of 2HGare measured prior to treatment, wherein an elevated level is indicatedfor the use of COMPOUND 1 or COMPOUND 2. Once the elevated levels areestablished, the level of 2HG is determined during the course of and/orfollowing termination of treatment to establish efficacy. In certainembodiments, the level of 2HG is only determined during the course ofand/or following termination of treatment. A reduction of 2HG levelsduring the course of treatment and following treatment is indicative ofefficacy. Similarly, a determination that 2HG levels are not elevatedduring the course of or following treatment is also indicative ofefficacy. Typically, 2HG measurements are utilized together with otherwell-known determinations of efficacy of malignancy treatment, such asreduction in number and size of tumors and/or other cancer-associatedlesions, improvement in the general health of the subject, andalterations in other biomarkers that are associated with malignancytreatment efficacy.

2HG can be detected in a sample by the methods of PCT Publication No. WO2011/050210 and US Publication No. US2012/0121515 hereby incorporated byreference in their entirety, or by analogous methods. In an exemplarymethod, 2HG can be detected in a sample by LC/MS. The sample is mixed80:20 with methanol, and centrifuged at 3,000 rpm for 20 minutes at 4degrees Celsius. The resulting supernatant can be collected and storedat −80 degrees Celsius prior to LC-MS/MS to assess 2-hydroxyglutaratelevels. A variety of different liquid chromatography (LC) separationmethods can be used. Each method can be coupled by negative electrosprayionization (ESI, −3.0 kV) to triple-quadrupole mass spectrometersoperating in multiple reaction monitoring (MRM) mode, with MS parametersoptimized on infused metabolite standard solutions. Metabolites can beseparated by reversed phase chromatography using 10 mM tributyl-amine asan ion pairing agent in the aqueous mobile phase, according to a variantof a previously reported method (Luo et al. J Chromatogr A 1147, 153-64,2007). One method allows resolution of TCA metabolites: t=0, 50% B; t=5,95% B; t=7, 95% B; t=8, 0% B, where B refers to an organic mobile phaseof 100% methanol. Another method is specific for 2-hydroxyglutarate,running a fast linear gradient from 50%-95% B (buffers as defined above)over 5 minutes. A Synergi Hydro-RP, 100 mm×2 mm, 2.1 μm particle size(Phenomonex) can be used as the column, as described above. Metabolitescan be quantified by comparison of peak areas with pure metabolitestandards at known concentration. Metabolite flux studies from¹³C-glutamine can be performed as described, e.g., in Munger et al. NatBiotechnol 26, 1179-86, 2008.

In one embodiment, 2HG is directly evaluated.

In another embodiment, a derivative of 2HG formed in process ofperforming the analytic method is evaluated. By way of example such aderivative can be a derivative formed in MS analysis. Derivatives caninclude a salt adduct, e.g., a Na adduct, a hydration variant, or ahydration variant which is also a salt adduct, e.g., a Na adduct, e.g.,as formed in MS analysis.

In another embodiment a metabolic derivative of 2HG is evaluated.Examples include species that build up or are elevated, or reduced, as aresult of the presence of 2HG, such as glutarate or glutamate that willbe correlated to 2HG, e.g., R-2HG.

Exemplary 2HG derivatives include dehydrated derivatives such as thecompounds provided below or a salt adduct thereof:

2HG is known to accumulate in the inherited metabolic disorder2-hydroxyglutaric aciduria. This disease is caused by deficiency in theenzyme 2-hydroxyglutarate dehydrogenase, which converts 2HG to α-KG(Struys, E. A. et al. Am J Hum Genet 76, 358-60 (2005)). Patients with2-hydroxyglutarate dehydrogenase deficiencies accumulate 2HG in thebrain as assessed by MRI and CSF analysis, develop leukoencephalopathy,and have an increased risk of developing brain tumors (Aghili, M.,Zahedi, F. & Rafiee, J Neurooncol 91, 233-6 (2009); Kolker, S.,Mayatepek, E. & Hoffmann, G. F. Neuropediatrics 33, 225-31 (2002);Wajner, M., Latini, A., Wyse, A. T. & Dutra-Filho, C. S. J Inherit MetabDis 27, 427-48 (2004)). Furthermore, elevated brain levels of 2HG resultin increased ROS levels (Kolker, S. et al. Eur J Neurosci 16, 21-8(2002); Latini, A. et al. Eur J Neurosci 17, 2017-22 (2003)),potentially contributing to an increased risk of cancer. The ability of2HG to act as an NMDA receptor agonist may contribute to this effect(Kolker, S. et al. Eur J Neurosci 16, 21-8 (2002)). 2HG may also betoxic to cells by competitively inhibiting glutamate and/or αKGutilizing enzymes. These include transaminases which allow utilizationof glutamate nitrogen for amino and nucleic acid biosynthesis, andαKG-dependent prolyl hydroxylases such as those which regulateHif1-alpha levels.

Treatment methods described herein can additionally comprise variousevaluation steps prior to and/or following treatment with COMPOUND 1 orCOMPOUND 2.

In one embodiment, prior to and/or after treatment with COMPOUND 1 orCOMPOUND 2, alone or in combination with a FLT3 pathway inhibitor, themethod further comprises the step of evaluating the growth, size,weight, invasiveness, stage and/or other phenotype of the malignancy.

In one embodiment, prior to and/or after treatment with COMPOUND 1,alone or in combination with a FLT3 pathway inhibitor, the methodfurther comprises the step of evaluating the IDH2 genotype of themalignancy. This may be achieved by ordinary methods in the art, such asDNA sequencing, immuno analysis, and/or evaluation of the presence,distribution or level of 2HG. In one embodiment, prior to and/or aftertreatment with COMPOUND 2, alone or in combination with a FLT3 pathwayinhibitor, the method further comprises the step of evaluating the IDH1genotype of the malignancy. This may be achieved by ordinary methods inthe art, such as DNA sequencing, immuno analysis, and/or evaluation ofthe presence, distribution or level of 2HG.

In one embodiment, prior to and/or after treatment with COMPOUND 1 orCOMPOUND 2, alone or in combination with a FLT3 pathway inhibitor, themethod further comprises the step of determining the 2HG level in thesubject. This may be achieved by spectroscopic analysis, e.g., magneticresonance-based analysis, e.g., MRI and/or MRS measurement, sampleanalysis of bodily fluid, such as serum or spinal cord fluid analysis,or by analysis of surgical material, e.g., by mass-spectroscopy.

In one embodiment, COMPOUND 1 and a FLT3 pathway inhibitor areadministered concurrently. In one embodiment, COMPOUND 1 and a FLT3pathway inhibitor are administered sequentially. In one embodiment,COMPOUND 2 and a FLT3 pathway inhibitor are administered concurrently.In one embodiment, COMPOUND 2 and a FLT3 pathway inhibitor areadministered sequentially.

In one embodiment, depending on the disease to be treated and thesubject's condition, COMPOUND 1 may be administered by oral, parenteral(e.g., intramuscular, intraperitoneal, intravenous, CIV, intracistemalinjection or infusion, subcutaneous injection, or implant), inhalation,nasal, vaginal, rectal, sublingual, or topical (e.g., transdermal orlocal) routes of administration. COMPOUND 1 may be formulated alone ortogether with one or more active agent(s), in suitable dosage unit withpharmaceutically acceptable excipients, carriers, adjuvants andvehicles, appropriate for each route of administration.

In one embodiment, depending on the disease to be treated and thesubject's condition, COMPOUND 2 may be administered by oral, parenteral(e.g., intramuscular, intraperitoneal, intravenous, CIV, intracistemalinjection or infusion, subcutaneous injection, or implant), inhalation,nasal, vaginal, rectal, sublingual, or topical (e.g., transdermal orlocal) routes of administration. COMPOUND 2 may be formulated alone ortogether with one or more active agent(s), in suitable dosage unit withpharmaceutically acceptable excipients, carriers, adjuvants andvehicles, appropriate for each route of administration.

In one embodiment, the amount of COMPOUND 1 or COMPOUND 2 administeredin the methods provided herein may range, e.g., between about 5 mg/dayand about 2,000 mg/day. In one embodiment, the range is between about 10mg/day and about 2,000 mg/day. In one embodiment, the range is betweenabout 20 mg/day and about 2,000 mg/day. In one embodiment, the range isbetween about 50 mg/day and about 1,000 mg/day. In one embodiment, therange is between about 100 mg/day and about 1,000 mg/day. In oneembodiment, the range is between about 100 mg/day and about 500 mg/day.In one embodiment, the range is between about 150 mg/day and about 500mg/day. In one embodiment, the range is or between about 150 mg/day andabout 250 mg/day. In certain embodiments, particular dosages are, e.g.,about 10 mg/day. In one embodiment, the dose is about 20 mg/day. In oneembodiment, the dose is about 50 mg/day. In one embodiment, the dose isabout 75 mg/day. In one embodiment, the dose is about 100 mg/day. In oneembodiment, the dose is about 120 mg/day. In one embodiment, the dose isabout 150 mg/day. In one embodiment, the dose is about 200 mg/day. Inone embodiment, the dose is about 250 mg/day. In one embodiment, thedose is about 300 mg/day. In one embodiment, the dose is about 350mg/day. In one embodiment, the dose is about 400 mg/day. In oneembodiment, the dose is about 450 mg/day. In one embodiment, the dose isabout 500 mg/day. In one embodiment, the dose is about 600 mg/day. Inone embodiment, the dose is about 700 mg/day. In one embodiment, thedose is about 800 mg/day. In one embodiment, the dose is about 900mg/day. In one embodiment, the dose is about 1,000 mg/day. In oneembodiment, the dose is about 1,200 mg/day. In one embodiment, the doseis or about 1,500 mg/day. In certain embodiments, particular dosagesare, e.g., up to about 10 mg/day. In one embodiment, the particular doseis up to about 20 mg/day. In one embodiment, the particular dose is upto about 50 mg/day. In one embodiment, the particular dose is up toabout 75 mg/day. In one embodiment, the particular dose is up to about100 mg/day. In one embodiment, the particular dose is up to about 120mg/day. In one embodiment, the particular dose is up to about 150mg/day. In one embodiment, the particular dose is up to about 200mg/day. In one embodiment, the particular dose is up to about 250mg/day. In one embodiment, the particular dose is up to about 300mg/day. In one embodiment, the particular dose is up to about 350mg/day. In one embodiment, the particular dose is up to about 400mg/day. In one embodiment, the particular dose is up to about 450mg/day. In one embodiment, the particular dose is up to about 500mg/day. In one embodiment, the particular dose is up to about 600mg/day. In one embodiment, the particular dose is up to about 700mg/day. In one embodiment, the particular dose is up to about 800mg/day. In one embodiment, the particular dose is up to about 900mg/day. In one embodiment, the particular dose is up to about 1,000mg/day. In one embodiment, the particular dose is up to about 1,200mg/day. In one embodiment, the particular dose is up to about 1,500mg/day.

In one embodiment, the amount of COMPOUND 1 or COMPOUND 2 in thepharmaceutical composition or dosage form provided herein may range,e.g., between about 5 mg and about 2,000 mg. In one embodiment, therange is between about 10 mg and about 2,000 mg. In one embodiment, therange is between about 20 mg and about 2,000 mg. In one embodiment, therange is between about 50 mg and about 1,000 mg. In one embodiment, therange is between about 50 mg and about 500 mg. In one embodiment, therange is between about 50 mg and about 250 mg. In one embodiment, therange is between about 100 mg and about 500 mg. In one embodiment, therange is between about 150 mg and about 500 mg. In one embodiment, therange is between about 150 mg and about 250 mg. In certain embodiments,particular amounts are, e.g., about 10 mg. In one embodiment, theparticular amount is about 20 mg. In one embodiment, the particularamount is about 30 mg. In one embodiment, the particular amount is about50 mg. In one embodiment, the particular amount is about 75 mg. In oneembodiment, the particular amount is about 100 mg. In one embodiment,the particular amount is about 120 mg. In one embodiment, the particularamount is about 150 mg. In one embodiment, the particular amount isabout 200 mg. In one embodiment, the particular amount is about 250 mg.In one embodiment, the particular amount is about 300 mg. In oneembodiment, the particular amount is about 350 mg. In one embodiment,the particular amount is about 400 mg. In one embodiment, the particularamount is about 450 mg. In one embodiment, the particular amount isabout 500 mg. In one embodiment, the particular amount is about 600 mg.In one embodiment, the particular amount is about 650 mg. In oneembodiment, the particular amount is about 700 mg. In one embodiment,the particular amount is about 800 mg. In one embodiment, the particularamount is about 900 mg. In one embodiment, the particular amount isabout 1,000 mg. In one embodiment, the particular amount is about 1,200mg. In one embodiment, the particular amount is or about 1,500 mg. Incertain embodiments, particular amounts are, e.g., up to about 10 mg. Inone embodiment, the particular amount is up to about 20 mg. In oneembodiment, the particular amount is up to about 50 mg. In oneembodiment, the particular amount is up to about 75 mg. In oneembodiment, the particular amount is up to about 100 mg. In oneembodiment, the particular amount is up to about 120 mg. In oneembodiment, the particular amount is up to about 150 mg. In oneembodiment, the particular amount is up to about 200 mg. In oneembodiment, the particular amount is up to about 250 mg. In oneembodiment, the particular amount is up to about 300 mg. In oneembodiment, the particular amount is up to about 350 mg. In oneembodiment, the particular amount is up to about 400 mg. In oneembodiment, the particular amount is up to about 450 mg. In oneembodiment, the particular amount is up to about 500 mg. In oneembodiment, the particular amount is up to about 600 mg. In oneembodiment, the particular amount is up to about 700 mg. In oneembodiment, the particular amount is up to about 800 mg. In oneembodiment, the particular amount is up to about 900 mg. In oneembodiment, the particular amount is up to about 1,000 mg. In oneembodiment, the particular amount is up to about 1,200 mg. In oneembodiment, the particular amount is up to about 1,500 mg.

In one embodiment, COMPOUND 1 or COMPOUND 2 can be delivered as a singledose such as, e.g., a single bolus injection, or oral tablets or pills;or over time such as, e.g., continuous infusion over time or dividedbolus doses over time. In one embodiment, compound 1 can be administeredrepetitively if necessary, for example, until the patient experiencesstable disease or regression, or until the patient experiences diseaseprogression or unacceptable toxicity. Stable disease or lack thereof isdetermined by methods known in the art such as evaluation of patient'ssymptoms, physical examination, visualization of the tumor that has beenimaged using X-ray, CAT, PET, or MRI scan and other commonly acceptedevaluation modalities.

In certain embodiments, COMPOUND 1 or COMPOUND 2 is administered to apatient in cycles (e.g., daily administration for one week, then a restperiod with no administration for up to three weeks). Cycling therapyinvolves the administration of an active agent for a period of time,followed by a rest for a period of time, and repeating this sequentialadministration. Cycling therapy can reduce the development ofresistance, avoid or reduce the side effects, and/or improves theefficacy of the treatment.

In one embodiment, a method provided herein comprises administeringCOMPOUND 1 or COMPOUND 2 in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, or greater than 40 cycles. In oneembodiment, the median number of cycles administered in a group ofpatients is about 1. In one embodiment, the median number of cyclesadministered in a group of patients is about 2. In one embodiment, themedian number of cycles administered in a group of patients is about 3.In one embodiment, the median number of cycles administered in a groupof patients is about 4. In one embodiment, the median number of cyclesadministered in a group of patients is about 5. In one embodiment, themedian number of cycles administered in a group of patients is about 6.In one embodiment, the median number of cycles administered in a groupof patients is about 7. In one embodiment, the median number of cyclesadministered in a group of patients is about 8. In one embodiment, themedian number of cycles administered in a group of patients is about 9.In one embodiment, the median number of cycles administered in a groupof patients is about 10. In one embodiment, the median number of cyclesadministered in a group of patients is about 11. In one embodiment, themedian number of cycles administered in a group of patients is about 12.In one embodiment, the median number of cycles administered in a groupof patients is about 13. In one embodiment, the median number of cyclesadministered in a group of patients is about 14. In one embodiment, themedian number of cycles administered in a group of patients is about 15.In one embodiment, the median number of cycles administered in a groupof patients is about 16. In one embodiment, the median number of cyclesadministered in a group of patients is about 17. In one embodiment, themedian number of cycles administered in a group of patients is about 18.In one embodiment, the median number of cycles administered in a groupof patients is about 19. In one embodiment, the median number of cyclesadministered in a group of patients is about 20. In one embodiment, themedian number of cycles administered in a group of patients is about 21.In one embodiment, the median number of cycles administered in a groupof patients is about 22. In one embodiment, the median number of cyclesadministered in a group of patients is about 23. In one embodiment, themedian number of cycles administered in a group of patients is about 24.In one embodiment, the median number of cycles administered in a groupof patients is about 25. In one embodiment, the median number of cyclesadministered in a group of patients is about 26. In one embodiment, themedian number of cycles administered in a group of patients is about 27.In one embodiment, the median number of cycles administered in a groupof patients is about 28. In one embodiment, the median number of cyclesadministered in a group of patients is about 29. In one embodiment, themedian number of cycles administered in a group of patients is about 30.In one embodiment, the median number of cycles administered in a groupof patients is greater than about 30 cycles.

In certain embodiments, treatment cycles comprise multiple doses ofCOMPOUND 1 or COMPOUND 2 administered to a subject in need thereof overmultiple days (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, orgreater than 14 days), optionally followed by treatment dosing holidays(e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or greater than 28 days).

In one embodiment, depending on the disease to be treated and thesubject's condition, the FLT3 pathway inhibitor may be administered byoral, parenteral (e.g., intramuscular, intraperitoneal, intravenous,CIV, intracistemal injection or infusion, subcutaneous injection, orimplant), inhalation, nasal, vaginal, rectal, sublingual, or topical(e.g., transdermal or local) routes of administration. In oneembodiment, the FLT3 pathway inhibitor may be formulated, alone ortogether with COMPOUND 1 and/or one or more active agent(s), in suitabledosage unit with pharmaceutically acceptable excipients, carriers,adjuvants and vehicles, appropriate for each route of administration. Inone embodiment, the FLT3 pathway inhibitor may be formulated, alone ortogether with COMPOUND 2 and/or one or more active agent(s), in suitabledosage unit with pharmaceutically acceptable excipients, carriers,adjuvants and vehicles, appropriate for each route of administration.

In one embodiment, the FLT3 pathway inhibitor is administered by, e.g.,intravenous (IV), subcutaneous (SC) or oral routes. Certain embodimentsherein provide co-administration of the FLT3 pathway inhibitor withCOMPOUND 1 or COMPOUND 2 and/or one or more additional active agents toprovide a synergistic therapeutic effect in subjects in need thereof.The co-administered active agent(s) may be cancer therapeutic agents, asdescribed herein. In certain embodiments, the co-administered activeagent(s) may be inhibitors of IDH1. In certain embodiments, theco-administered active agent(s) may be inhibitors of IDH2. In certainembodiments, the co-administered agent(s) may be dosed, e.g., orally orby injection (e.g., IV or SC).

In certain embodiments, treatment cycles comprise multiple doses of theFLT3 pathway inhibitor administered to a subject in need thereof overmultiple days (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, orgreater than 14 days), optionally followed by treatment dosing holidays(e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or greater than 28 days).Suitable dosage amounts for the methods provided herein include, e.g.,therapeutically effective amounts and prophylactically effectiveamounts.

In one embodiment, the FLT3 pathway inhibitor can be delivered as asingle dose such as, e.g., a single bolus injection, or oral tablets orpills; or over time such as, e.g., continuous infusion over time ordivided bolus doses over time. In one embodiment, the FLT3 pathwayinhibitor can be administered repetitively if necessary, for example,until the patient experiences stable disease or regression, or until thepatient experiences disease progression or unacceptable toxicity. Stabledisease or lack thereof is determined by methods known in the art suchas evaluation of patient's symptoms, physical examination, visualizationof the tumor that has been imaged using X-ray, CAT, PET, or MRI scan andother commonly accepted evaluation modalities.

In one embodiment, the FLT3 pathway inhibitor can be administered oncedaily or divided into multiple daily doses such as twice daily, threetimes daily, and four times daily. In one embodiment, the administrationcan be continuous (i.e., daily for consecutive days or every day),intermittent, e.g., in cycles (i.e., including days, weeks, or months ofrest when no drug is administered). In one embodiment, the FLT3 pathwayinhibitor is administered daily, for example, once or more than onceeach day for a period of time. In one embodiment, the FLT3 pathwayinhibitor is administered intermittently, i.e., stopping and starting ateither regular or irregular intervals.

Patient Population

In certain embodiments of the methods provided herein, the subject to betreated is an animal, for example a mammal or a non-human primate. Inparticular embodiments, the subject is a human patient. The subject canbe male or female.

Particularly, subjects amenable to treatment according to the methodsprovided herein include subjects with cancer, wherein the cancer ischaracterized by the presence of a mutant allele of IDH1 and/or IDH2 andthe absence of a FLT3 mutation.

In certain embodiments, subjects amenable to treatment according to themethods provided herein include subjects with cancer, wherein the canceris characterized by the presence of a mutant allele of IDH1 and/or IDH2and further characterized by a mutant FLT3.

In one embodiment, subjects amenable to treatment according to themethods provided herein include subjects with advanced hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS, myeloproliferative neoplasms (MPN), chronicmyelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias (B-ALL),or lymphoma (e.g., T-cell lymphoma), each characterized by the presenceof a mutant allele of IDH1 and the absence of a FLT3 mutation.

In one embodiment, subjects amenable to treatment according to themethods provided herein include subjects with advanced hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), chronic myelomonocytic leukemia (CMML), myeloid sarcoma,multiple myeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, each characterized by the presence of a mutantallele of IDH2 and the absence of a FLT3 mutation.

In one embodiment, subjects amenable to treatment according to themethods provided herein include subjects with a solid tumors, such asglioma, melanoma, chondrosarcoma, cholangiocarcinoma (includingintrahepatic cholangiocarcinoma (IHCC), prostate cancer, colon cancer,or non-small cell lung cancer (NSCLC), each characterized by thepresence of a mutant allele of IDH1, and the absence of a FLT3 mutation.

In one embodiment, subjects amenable to treatment according to themethods provided herein include subjects with a solid tumor, such asglioma, melanoma, chondrosarcoma, cholangiocarcinoma (e.g., glioma),angioimmunoblastic T-cell lymphoma (AITL), sarcoma, or non small celllung cancer, each characterized by the presence of a mutant allele ofIDH2 and the absence of a FLT3 mutation.

In certain embodiments, subjects amenable to treatment according to themethods provided herein include subjects with advanced hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS, myeloproliferative neoplasms (MPN), chronicmyelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias (B-ALL),or lymphoma (e.g., T-cell lymphoma), each characterized by the presenceof a mutant allele of IDH1 and further characterized by one or more FLT3mutation(s).

In certain embodiments, subjects amenable to treatment according to themethods provided herein include subjects with advanced hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), chronic myelomonocytic leukemia (CMML), myeloid sarcoma,multiple myeloma, lymphoma (e.g., T-cell lymphoma or B-cell lymphoma),angioimmunoblastic T-cell lymphoma (AITL) or blastic plasmacytoiddendritic cell neoplasm, each characterized by the presence of a mutantallele of IDH2 and further characterized by one or more FLT3mutation(s).

In certain embodiments, subjects amenable to treatment according to themethods provided herein include subjects with a solid tumors, such asglioma, melanoma, chondrosarcoma, cholangiocarcinoma (includingintrahepatic cholangiocarcinoma (IHCC), prostate cancer, colon cancer,or non-small cell lung cancer (NSCLC), each characterized by thepresence of a mutant allele of IDH1 and further characterized by one ormore FLT3 mutation(s).

In certain embodiments, subjects amenable to treatment according to themethods provided herein include subjects with a solid tumor, such asglioma, melanoma, chondrosarcoma, cholangiocarcinoma (e.g., glioma),angioimmunoblastic T-cell lymphoma (AITL), sarcoma, or non small celllung cancer, each characterized by the presence of a mutant allele ofIDH2 and further characterized by one or more FLT3 mutation(s).

Also encompassed are methods of treating a subject regardless of thesubject's age, although some diseases or disorders are more common incertain age groups. In some embodiments, the subject is a human patientat least 18 years old. In some embodiments, the patient is 10, 15, 18,21, 24, 35, 40, 45, 50, 55, 65, 70, 75, 80, or 85 years old or older.

In certain embodiments, the methods provided herein encompass thetreatment of subjects who have not been previously treated for cancer.In other embodiments, the methods encompass treating subjects who havebeen previously treated but are non-responsive to standard therapies aswell as those who are currently being treated for cancer. For example,the subjects may have been previously treated or are currently beingtreated with a standard treatment regimen for cancer known to thepractitioner of skill in the art.

It is understood that the foregoing detailed description andaccompanying examples are merely illustrative, and are not to be takenas limitations upon the scope of the subject matter. Various changes andmodifications to the disclosed embodiments will be apparent to thoseskilled in the art. Such changes and modifications, including withoutlimitation those relating to the methods of use provided herein, may bemade without departing from the spirit and scope thereof. Patents,patent publications, and other publications referenced herein areincorporated by reference.

EXAMPLES

As used herein, the symbols and conventions used in the examples,regardless of whether a particular abbreviation is specifically defined,are consistent with those used in the contemporary scientificliterature, for example, the Journal of the American Chemical Society orthe Journal of Biological Chemistry. Specifically, but withoutlimitation, the following abbreviations may be used in the examples andthroughout the specification: FMI=Foundation Medicine Inc.; CR=completeremission; CRi=complete remision with incomplete blood count recovery;CRp=complete remision with incomplete platelet recovery;MLFS=morphologic leukemia-free state; FLT3-ITD−=Fms Related TyrosineKinase 3 internal tandem repeat negative; FLT3-ITD+=Fms Related TyrosineKinase 3 internal tandem repeat positive; mFLT3=mutant Fms RelatedTyrosine Kinase 3 internal tandem repeat; ORR=overall response rate;PM−=point mutation negative; PM+=point mutation positive; g (grams); mg(milligrams); mL (milliliters); mL (microliters); M (molar); mM(millimolar); mM (micromolar); hr or hrs (hour or hours); and min(minutes).

Example 1. A Phase 1/2, Multicenter, Open-Label, Dose-Escalation andExpansion, Safety, Pharmacokinetic, Pharmacodynamic, and ClinicalActivity Study of Orally Administered COMPOUND 1 in Subjects withAdvanced Hematologic Malignancies with an IDH2 Mutation

Indication: Treatment of patients with advanced hematologic malignancieswith an IDH2 mutation.

Phase 1 (Dose Escalation and Part 1 Expansion) Objectives:

Primary Objectives

To assess the safety and tolerability of treatment with COMPOUND 1administered continuously as a single agent dosed orally on Days 1 to 28of a 28-day cycle in subjects with advanced hematologic malignancies.

To determine a maximum tolerated dose (MTD) or a maximum administereddose (MAD) and/or the recommended Phase 2 dose (RP2D) of COMPOUND 1 insubjects with advanced hematologic malignancies.

Secondary Objectives

To describe the dose-limiting toxicities (DLTs) of COMPOUND 1 insubjects with advanced hematologic malignancies.

To characterize the pharmacokinetics (PK) of COMPOUND 1 and itsmetabolite in subjects with advanced hematologic malignancies.

To characterize the PK/pharmacodynamic (PD) relationship of COMPOUND 1and 2-hydroxygluturate (2-HG).

To characterize the clinical activity associated with COMPOUND 1 insubjects with advanced hematologic malignancies.

Phase 2 Objectives:

Primary Objectives

To assess the efficacy of COMPOUND 1 as treatment for subjects withrelapsed or refractory AML with an IDH2 mutation.

Secondary Objectives

To further evaluate the safety profile of COMPOUND 1 in subjects withrelapsed or refractory AML with an IDH2 mutation.

To characterize the pharmacokinetics (PK) of COMPOUND 1 and itsmetabolite in subjects with relapsed or refractory AML with an IDH2mutation.

To characterize the PK/pharmacodynamic (PD) relationship of COMPOUND 1and 2-hydroxygluturate (2-HG).

Methodology:

This is a Phase 1/2, multicenter, open-label, 3-part (Phase 1 doseescalation, Phase 1 Part 1 Expansion, and Phase 2), safety, PK/PD, andclinical activity evaluation of orally administered COMPOUND 1 insubjects with advanced hematologic malignancies with an IDH2 mutation.

In the Phase 1 portion, the study includes a dose escalation phase todetermine MTD/MAD and/or the RP2D and an expansion phase (Part 1Expansion) to further evaluate the safety, tolerability and clinicalactivity of COMPOUND 1 in select populations. The Phase 2 portion(previously Part 2 Expansion) will further inform on the efficacy,safety, tolerability and clinical activity of COMPOUND 1 in subjectswith refractory or relapsed AML with an IDH2 mutation.

Dose Escalation Phase

The dose escalation phase will utilize a standard “3+3” design. Duringthe dose escalation phase, consented eligible subjects with relapsed orrefractory acute myelogenous leukemia (AML), untreated AML≥60 years ofage who are not candidates for standard therapy, or myelodysplasticsyndrome with refractory anemia with excess blasts will be enrolled intosequential cohorts of increasing doses of AG-221 not to exceed 650 mg QDdose. Each dose cohort will enroll a minimum of 3 subjects. The first 3subjects enrolled in each dosing cohort during the dose escalationportion of the study will receive a single dose of study drug on Day −3(i.e., 3 days prior to the start of daily dosing) and undergo safety andPK/PD assessments over 72 hours to evaluate drug concentrations and 2-HGand α-KG levels. The next dose of study drug will be on Cycle 1 Day 1(C1D1) at which time daily dosing will begin. The initial dosingschedule was twice daily (approximately every 12 hours). Based on theemerging data, a once daily dosing schedule also has been implemented.Alternative dosing schedules (e.g., a loading dose followed by oncedaily dosing) may continue to be explored in the dose escalation andexpansion phases as agreed upon by the Clinical Study Team. If there aremultiple subjects in the screening process at the time the third subjectwithin a cohort begins treatment, up to 2 additional subjects may beenrolled with approval of the Medical Monitor. For these additionalsubjects, the Day −3 through Day 1 PK/PD assessments are optionalfollowing discussion with the Medical Monitor.

The safety of dosing during the dose escalation phase will be evaluatedby the Clinical Study Team, comprised of the Sponsor designee(Responsible Medical Officer), Study Medical Monitor, and Investigators.The Clinical Study Team will review the emerging safety data from eachcohort to determine if dose escalation will occur.

Toxicity severity will be graded according to the National CancerInstitute Common Terminology Criteria for Adverse Events (NCI CTCAE)Version 4.03. A DLT is defined as outlined below.

Non-Hematologic:

All clinically significant non-hematologic toxicities CTCAE≥Grade 3 withthe exception of ≥Grade 3 blood bilirubin increases in subjects with aUDP (uridine diphosphate)-glucuronosyltransferase 1 family, polypeptideA1 (UGT1A1) mutation. In subjects with a UGT1A1 mutation, bloodbilirubin increases of >5× upper limit of normal (ULN) may be considereda DLT.

Hematologic:

Prolonged myelosuppression, defined as persistence of ≥Grade 3neutropenia or thrombocytopenia (by NCI CTCAE, version 4.03,leukemia-specific criteria, i.e., marrow cellularity <5% on Day 28 orlater from the start of study drug without evidence of leukemia) atleast 42 days after the initiation of Cycle 1 therapy. Leukemia-specificgrading should be used for cytopenias (based on percentage decrease frombaseline: 50 to 75%=Grade 3, >75%=Grade 4).

Due to frequent co-morbidities and concurrent medications in thepopulation under study, attribution of adverse events (AEs) to aparticular drug is challenging. Therefore, all AEs that cannot clearlybe determined to be unrelated to COMPOUND 1 will be considered relevantto determining DLTs and will be reviewed by the Clinical Study Team. TheClinical Study Team also will review any other emergent toxicities thatare not explicitly defined by the DLT criteria to determine if anywarrant a DLT designation.

If, after the third subject completes the 28-day DLT evaluation period(i.e., Cycle 1), no DLTs are observed, the study will proceed with doseescalation to the next cohort following safety review by the ClinicalStudy Team. If 1 of 3 subjects experiences a DLT during the first cycle,3 additional subjects will be enrolled in that cohort. If none of theadditional 3 subjects experience a DLT, dose escalation may continue tothe next cohort following safety review by the Clinical Study Team. If 2or more subjects in a cohort experience DLTs during the first cycle,dose escalation will be halted and the next lower dose level will bedeclared the MTD. If the MTD cohort included only 3 subjects, anadditional 3 subjects will be enrolled at that dose level to confirmthat <2 of 6 subjects experience a DLT at that dose. Alternatively, adose level intermediate between the non-tolerated dose level and thepreviously tolerated dose level may be explored and declared the MTD if<2 out of 6 subjects experience a DLT at that dose.

Increases in the dose of AG-221 for each dose cohort will be guided byan accelerated titration design, where the dose will be doubled (100%increase) from one cohort to the next until AG-221-related NCI CTCAEGrade 2 or greater toxicity is observed in any subject within thecohort. Following evaluation by the Clinical Study Team, subsequentincreases in dose will be 50% or less until the MTD is determined. Theabsolute percent increase in the dose will be determined by the ClinicalStudy Team predicated on the type and severity of any toxicity seen inthe prior dose cohorts. The MTD is the highest dose that causes DLTs in<2 of 6 subjects.

To optimize the number of subjects treated at a potentially clinicallyrelevant dose, intra-subject dose escalation will be permitted withapproval of the Medical Monitor.

Part 1 Expansion Phase

During the Part 1 expansion phase, safety, PK/PD, and preliminaryclinical activity data will be reviewed by the Clinical Study Team on anongoing basis.

In Part 1, 4 non-randomized cohorts of approximately 25 subjects per armwith IDH2-mutated hematologic malignancies will be enrolled as follows:

Arm 1: Relapsed or refractory AML and age ≥60 years, or any subject withAML regardless of age who has relapsed following a bone marrowtransplant (BMT).

Arm 2: Relapsed or refractory AML and age <60 years, excluding subjectswith AML who have relapsed following a BMT.

Arm 3: Untreated AML and age ≥60 years that decline standard of carechemotherapy.

Arm 4: IDH2-mutated advanced hematologic malignancies not eligible forArms 1 to 3.

Phase 2

Phase 2, the pivotal part of the study, will further establish theefficacy and safety profile of COMPOUND 1 at the recommended phase 2dose (RP2D) determined in the ongoing dose escalation phase in subjectswith IDH2-mutated relapsed or refractory AML defined as follows:

Subjects who relapse after allogeneic transplantation

Subjects in second or later relapse;

Subjects who are refractory to initial induction or re-inductiontreatment;

Subjects who relapse within 1 year of initial treatment, excludingpatients with favorable-risk status according to NCCN Guidelines.Favorable-risk cytogenetics: inv(16), +(16;16), t(8;21), t(15;17).

Approximately 125 subjects will be enrolled in this part of the trial.

General Study Conduct:

Following informed consent, all subjects will undergo screeningprocedures within 28 days prior to the C1D1 to determine eligibility.All subjects are required to have confirmation of IDH2-mutated diseasefrom a bone marrow aspirate and peripheral blood. For subjects in thedose escalation phase and Part 1 Expansion, documentation of IDH2mutation status can be based on local site testing with centrallaboratory testing performed retrospectively. Subjects in Phase 2 arerequired to have IDH2-mutation status based on central laboratorytesting during screening prior to study treatment. Additional screeningprocedures include medical, surgical, and medication history, a buccalswab for germ-line mutation analysis, physical examination, vital signs,Eastern Cooperative Oncology Group (ECOG) performance status (PS),12-lead electrocardiogram (ECG), evaluation of left ventricular ejectionfraction (LVEF), clinical laboratory assessments (hematology, chemistry,coagulation, and serum pregnancy test), bone marrow biopsy and aspirate,blood and bone marrow samples for 2-HG and α-KG measurement, and bloodfor determination of UGT1A1 mutation status. In addition, subjects inthe Part 1 Expansion will have urine samples for 2-HG and α-KGmeasurement and blood samples for cholesterol, and 4β-0H-cholesterollevels collected during screening.

Dose Escalation and Part 1 Expansion

Three days prior to the start of daily dosing of AG-221 (Day −3), thefirst 3 subjects enrolled in each cohort in the dose escalation phaseand the first 15 subjects enrolled in each arm of Part 1 Expansion willreceive a single dose of AG-221 in clinic and have serial blood andurine samples obtained for determination of blood and urineconcentrations of AG-221, its metabolite AGI-16903, 2-HG, and α-KG. Afull 72-hour PK/PD profile will be conducted: subjects will be requiredto remain at the study site for 10 hours on Day −3 and return on Days−2, −1, and 1 for 24-, 48-, and 72-hour samples, respectively. Duringthe in-clinic period on Day −3, clinical observation and serial 12-leadECGs and vital signs assessments will be conducted. Daily treatment withAG-221 will begin on C1D1; for subjects in the dose escalation phase andPart 1 Expansion who did not undergo the Day −3 PK/PD assessments,clinical observation and serial 12-lead ECGs and vital signs assessmentswill be conducted over 8 hours following their first dose of AG-221 onC1D1.

Subjects in the dose escalation phase and Part 1 Expansion also willundergo PK/PD assessments over a 10-hour period on C1D15, C2D1, andC4D1. Predose blood samples (trough) will be obtained on C1D1 (for thosesubjects who did not undergo the Day −3 PK/PD assessments), C1D8, C1D22,C2D15, C3D1, C3D15, C5D1, and Day 1 of all cycles thereafter fordetermination of AG-221, 2-HG, and α-KG concentrations. These subjectswill have urine collected for PK/PD evaluation at screening; prior todosing on C1D15, C2D1 and Day 1 of all cycles thereafter; and at the Endof Treatment visit. Available bone marrow biopsy samples also will beassessed for 2-HG and α-KG levels.

Phase 2

Subjects in the Phase 2 portion of the trial are not required to undergothe Day 3 assessments; these subjects will undergo an 8-hour PK/PDprofile conducted on Day 1 of Cycles 1 and 2, and predose blood samples(trough) on C1D2 and C2D2 will be obtained in order to assess PK/PD in a24-hour period. Additional blood samples for PK/PD assessments will bedrawn pre-dose (within 30 minutes) on Day 1 of Cycle 3, and at the Endof Treatment visit. Time-matched 12-lead ECGs will be conducted intriplicate on Day 1 of Cycles 1 and 2; a triplicate ECG is also to beobtained at the End of Treatment visit. Single 12-lead ECGs will beconducted on Day 1 of every cycle beginning with Cycle 3, and at theFollow-up visit. Available bone marrow biopsy samples will be assessedfor 2-HG and α-KG levels.

Other Safety Assessments (All Phases)

All subjects will undergo safety assessments during the treatment periodto include physical examination, vital signs, ECOG PS, 12-lead ECGs,evaluation of LVEF, and clinical laboratory assessments (hematology,chemistry, coagulation, and pregnancy testing).

Clinical Activity Assessments:

Phase 1 (Dose Escalation and Part 1 Expansion)

Subjects in the dose escalation phase and Part 1 Expansion will have theextent of their disease assessed, including bone marrow biopsies and/oraspirates and peripheral blood, at screening, on C1D15, C2D1, and C3D1,every 28 days (peripheral blood only) or every 56 days (bone marrowbiopsies and/or aspirates and peripheral blood) thereafter while onstudy drug treatment, independent of dose delays and/or doseinterruptions, and/or at any time when progression of disease issuspected. Response to treatment and treatment decisions in all subjectswill be determined by the Investigators based on modified InternationalWorking Group (IWG) response criteriaor other appropriate responsecriteria for the malignancy under study.

Phase 2

For subjects enrolled in the Phase 2 portion of the trial, extent ofdisease, including bone marrow biopsies and/or aspirates and peripheralblood, will be assessed at screening, on C2D1, every 28 days thereafterthrough 12 months, and every 56 days thereafter while on study drugtreatment, independent of dose delays and/or dose interruptions, and/orat any time when progression of disease is suspected. Eligibility,treatment decisions, and response to treatment will be determined by theInvestigators based on modified International Working Group (IWG)response criteria. Response will be also be assessed retrospectively byan Independent Response Adjudication Committee (IRAC).

End of Treatment and Follow-up:

Subjects may continue treatment with COMPOUND 1 until diseaseprogression or development of unacceptable toxicity.

Evidence supports that cancer-associated IDH mutations block normalcellular differentiation and promote tumorigenesis via the abnormalproduction of 2-HG, a potential oncometabolite. COMPOUND 1 may produceantitumor effects by reversing the differentiation block induced by theIDH2 mutations and promoting appropriate cellular differentiation.

Because of the unique mechanism of action of COMPOUND 1, clinicalresponses are different than those observed with cytotoxic agents.Responses with COMPOUND 1 may be occur after 2 or more cycles of therapyand they may occur after an initiation period of leukocytosis in theperipheral blood and/or bone marrow with, in rare cases, correspondingclinical signs and symptoms of fever, fluid retention, hypoxia, and skinrash which have been termed a differentiation-like syndrome.

As such, standard assessment criteria developed based on the experiencefrom the cytotoxic chemotherapeutic agents do not provide a complete andaccurate response assessment for this novel class of IDH2 inhibitors.Therefore, in the setting where a subject's assessment shows signssimilar to progression within the first 2 cycles, caution should beexercised in discontinuing study drug, and a discussion with the MedicalMonitor is required, especially in situations where the subject'sclinical condition is stable as supported by, but limited to, absence ofsigns and symptoms of rapid deterioration indicating disease progressionand/or general condition is stable or improving.

Subjects who experience progression of disease (PD) per the applicableresponse criteria, should have assessment of the disease repeated 28days later in order to confirm PD with option of continuing treatment asdescribed above while awaiting for confirmation. If repeat evaluationconfirms PD subjects will discontinue study treatment and proceed to thesurvival follow-up phase.

Subjects with stable or progressive disease may continue to receivestudy treatment with AG-221 at the discretion of the Investigator andwith Medical Monitor approval.

All subjects are to undergo an end of treatment assessment (withinapproximately 5 days of the last dose of study drug); in addition, afollow-up safety assessment is to be scheduled 28 days after the lastdose. Furthermore, all subjects will be followed monthly for diseasestatus, overall survival, and initiation of non-study anti-neoplastictherapy, until death, withdrawal of consent, or the end of the study,whichever occurs first.

Subjects who achieve an adequate response to treatment with AG-221 andmeet other criteria required to undergo hematopoietic stem celltransplant (HSCT) may proceed to HSCT after discontinuation of studytherapy. Those subjects will be followed on study for outcome untilrelapse or end of study to support the overall clinical benefit ofAG-221 in this setting.

Subjects who relapse following HSCT may be eligible to restart treatmentwith AG-221 with Medical Monitor approval and at the discretion of theInvestigator, if they have confirmed recurrent IDH2 mutant positivedisease, no other cancer treatment (with the exception ofanti-neoplastic therapies used in the course of HSCT such asconditioning regimen or induction-type regimen and anti-GVHD prophylaxis[i.e., methotrexate]) besides HSCT was administered since the last doseof AG-221, the subject meets the safety parameters listed in theInclusion/Exclusion criteria, and the trial is open. Subjects willresume AG-221 therapy at the same dose and schedule at the time ofAG-221 treatment discontinuation prior to HSCT.

All subjects, including those who relapse following HSCT and elect notto restart treatment, will be followed monthly thereafter for assessmentof survival status and non-study anti-neoplastic therapies sincediscontinuation of study drug until death or end of study.

Number of Subjects (Planned):

Approximately a minimum of 291 subjects in total is planned to beenrolled in the study (i.e., in the dose escalation, Part 1 Expansion,and Phase 2 portion of the trial).

Assuming that identification of the MTD/MAD requires the evaluation of13 dose levels/schedules of COMPOUND 1 with up to 5 subjects per doselevel, with the exception that the MTD/MAD requires 6 subjects, then 66subjects will be enrolled during the dose escalation part of the study.Additional subjects may be needed for cohort expansion during doseescalation, for the replacement of subjects who are not evaluable forPK/PD, safety, or clinical activity, or for evaluation of alternativedosing regimens other than the planned escalation scheme or the MTD/MAD,to optimize the RP2D and regimen(s). Dose levels (ranging from 30 mg to150 mg) have been evaluated in the BID schedule and 8 dose levels(ranging from 50 mg to 650 mg) have been evaluated in the QD schedule.

Four cohorts of a minimum of 25 additional subjects in specifichematologic malignancy subsets (total a minimum of 100 subjects) will beenrolled in Part 1 Expansion of the study.

The Phase 2 portion of the trial will enroll approximately 125 subjectswith relapsed or refractory AML with an IDH2 mutation. Additionalsubjects may be needed for the replacement of subjects who are notevaluable for PK/PD, safety, and/or clinical activity, or for evaluationof alternative dosing regimens. The final total sample size may beadjusted according to the observed toxicity rate, and number of subjectsenrolled for expanded evaluation.

Inclusion Criteria

Subjects must meet all of the following criteria to be enrolled in thestudy:

-   -   1. Subject must be ≥18 years of age.    -   2. Subjects must have advanced hematologic malignancy including:

Phase 1/Dose Escalation:

-   -   Diagnosis of AML according to World Health Organization (WHO)        criteria;        -   Disease refractory or relapsed (defined as the reappearance            of >5% blasts in the bone marrow).        -   Untreated AML, ≥60 years of age and are not candidates for            standard therapy due to age, performance status, and/or            adverse risk factors, according to the treating physician            and with approval of the Medical Monitor;    -   Diagnosis of MDS according to WHO classification with refractory        anemia with excess blasts (subtype RAEB-1 or RAEB-2), or        considered high-risk by the Revised International Prognostic        Scoring System (IPSS-R) that is recurrent or refractory, or the        subject is intolerant to established therapy known to provide        clinical benefit for their condition (i.e., subjects must not be        candidates for regimens known to provide clinical benefit),        according to the treating physician and with approval of the        Medical Monitor. (Subjects with other relapsed and/or primary        refractory hematologic cancers, for example CMML, who fulfill        the inclusion/excluding criteria may be considered on a case-by        case basis, with approval of the Medical Monitor.)

Phase 1/Part 1 Expansion:

-   -   Arm 1: Relapsed or refractory AML and age ≥60 years, or any        subject with AML regardless of age who has relapsed following a        BMT.    -   Arm 2: Relapsed or refractory AML and age <60 years, excluding        subjects with AML who have relapsed following a BMT.    -   Arm 3: Untreated AML and age ≥60 years that decline standard of        care chemotherapy.    -   Arm 4: IDH2-mutated advanced hematologic malignancies not        eligible for Arms 1 to 3.

Phase 2:

-   -   Diagnosis of AML according to World Health Organization (WHO)        criteria and disease relapsed or refractory as defined by:        -   Subjects who relapse after allogeneic transplantation;        -   Subjects in second or later relapse;        -   Subjects who are refractory to initial induction or            re-induction treatment;        -   Subjects who relapse within 1 year of initial treatment,            excluding patients with favorable-risk status according to            NCCN Guidelines. Favorable-risk cytogenetics: inv(16),            +(16;16), t(8;21), t(15;17).    -   3. Subjects must have documented IDH2 gene-mutated disease:    -   For subjects in the dose escalation phase and Part 1 Expansion,        IDH2 mutation may be based on local evaluation. (Centralized        testing will be performed retrospectively).    -   4. For subjects in the Phase 2 portion of the trial, central        testing of IDH2 mutation in samples of bone marrow aspirate and        peripheral blood, is required during screening to confirm        eligibility. Subjects must be amenable to serial bone marrow        sampling, peripheral blood sampling, and urine sampling during        the study.    -   The diagnosis and evaluation of AML or MDS will be made by bone        marrow aspiration and biopsy. If an aspirate is unobtainable        (i.e., a “dry tap”), the diagnosis may be made from the core        biopsy.    -   Screening bone marrow aspirate and peripheral blood samples are        required for all subjects. A bone marrow biopsy must be        collected if adequate aspirate is not attainable unless:        -   A bone marrow aspirate and biopsy was performed as part of            the standard of care within 28 days prior to the start of            the study treatment; and        -   Slides of bone marrow aspirate, biopsy and stained            peripheral blood smear are available for both local and            central pathology reviewers;    -   5. Subjects must be able to understand and willing to sign an        informed consent. A legally authorized representative may        consent on behalf of a subject who is otherwise unable to        provide informed consent, if acceptable to, and approved by, the        site and/or site's Institutional Review Board (IRB)/Independent        Ethic Committee (IEC).    -   6. Subjects must have ECOG PS of 0 to 2.    -   7. Platelet count ≥20,000/μL (Transfusions to achieve this level        are allowed.) Subjects with a baseline platelet count of        <20,000/μL due to underlying malignancy are eligible with        Medical Monitor approval.    -   8. Subjects must have adequate hepatic function as evidenced by:    -   Serum total bilirubin ≤1.5× upper limit of normal (ULN), unless        considered due to Gilbert's disease, a gene mutation in UGT1A1,        or leukemic organ involvement, following approval by the Medical        Monitor;    -   Aspartate aminotransferase (AST), alanine aminotransferase        (ALT), and alkaline phosphatase (ALP)≤3.0×ULN, unless considered        due to leukemic organ involvement.    -   9. Subjects must have adequate renal function as evidenced by:    -   Serum creatinine ≤2.0×ULN    -   OR    -   Creatinine clearance >40 mL/min based on the Cockroft-Gault        glomerular filtration rate (GFR) estimation:

(140−Age)×(weight in kg)×(0.85 if female)/72×serum creatinine

-   -   10. Subjects must be recovered from any clinically relevant        toxic effects of any prior surgery, radiotherapy, or other        therapy intended for the treatment of cancer. (Subjects with        residual Grade 1 toxicity, for example Grade 1 peripheral        neuropathy or residual alopecia, are allowed with approval of        the Medical Monitor.)    -   11. Female subjects with reproductive potential must agree to        undergo medically supervised pregnancy test prior to starting        study drug. The first pregnancy test will be performed at        screening (within 7 days prior to first study drug        administration), and on the day of the first study drug        administration and confirmed negative prior to dosing and Day 1        before dosing all subsequent cycles.    -   12. Female subjects with reproductive potential must have a        negative serum pregnancy test within 7 days prior to the start        of therapy. Subjects with reproductive potential are defined as        sexually mature women who have not undergone a hysterectomy,        bilateral oophorectomy or tubal occlusion or who have not been        naturally postmenopausal (i.e., who have not menstruated at all)        for at least 24 consecutive months (i.e., has had menses at any        time in the preceding 24 consecutive months). Females of        reproductive potential as well as fertile men and their partners        who are female of reproductive potential must agree to abstain        from sexual intercourse or to use two highly effective forms of        contraception from the time of giving informed consent, during        the study and for 120 days (females and males) following the        last dose of COMPOUND 1. A highly effective form of        contraception is defined as hormonal oral contraceptives,        injectables, patches, intrauterine devices, double-barrier        method (e.g., synthetic condoms, diaphragm, or cervical cap with        spermicidal foam, cream, or gel), or male partner sterilization.    -   13. Able to adhere to the study visit schedule (ie, clinic        visits at the study sites are mandatory, unless noted otherwise        for particular study visits) and other protocol requirements

Exclusion Criteria

Subjects who meet any of the following criteria will not be enrolled inthe study:

-   -   1. Subjects who have undergone a hematopoietic stem cell        transplant (HSCT) within 60 days of the first dose of COMPOUND        1, or subjects on immunosuppressive therapy post HSCT at the        time of screening, or with clinically significant        graft-versus-host disease (GVHD). (The use of a stable dose of        oral steroids post HSCT and/or topical steroids for ongoing skin        GVHD is permitted with Medical Monitor approval.)    -   2. Subjects who received systemic anticancer therapy or        radiotherapy <14 days prior to their first day of study drug        administration. (Hydroxyurea is allowed prior to enrollment and        after the start of COMPOUND 1 for the control of peripheral        leukemic blasts in subjects with leukocytosis (white blood cell        [WBC] counts >30,0004 μL).    -   3. Subjects who received a small molecule investigational agent        <14 days prior to their first day of study drug administration.        In addition, the first dose of COMPOUND 1 should not occur        before a period ≥5 half-lives of the investigational agent has        elapsed.    -   4. Subjects taking the following sensitive CYP substrate        medications that have a narrow therapeutic range are excluded        from the study unless they can be transferred to other        medications within ≥5 half-lives prior to dosing: paclitaxel        (CYP2C8) warfarin, phenytoin (CYP2C9), S-mephenytoin (CYP2C19),        thioridazine (CYP2D6), theophylline and tizanidine (CYP1A2).    -   5. Subjects taking the P-gp and BCRP transporter-sensitive        substrates digoxin and rosuvastatin should be excluded from the        study unless they can be transferred to other medications within        ≥5 half-lives prior to dosing.    -   6. Subjects for whom potentially curative anticancer therapy is        available.    -   7. Subjects who are pregnant or lactating.    -   8. Subjects with an active severe infection that required        anti-infective therapy or with an unexplained fever >38.5° C.        during screening visits or on their first day of study drug        administration (at the discretion of the Investigator, subjects        with tumor fever may be enrolled).    -   9. Subjects with known hypersensitivity to any of the components        of COMPOUND 1.    -   10. Subjects with New York Heart Association (NYHA) Class III or        IV congestive heart failure or LVEF <40% by echocardiogram        (ECHO) or multi-gated acquisition (MUGA) scan obtained within        approximately 28 days of C1D1.    -   11. Subjects with a history of myocardial infarction within the        last 6 months of screening.    -   12. Subjects with uncontrolled hypertension (systolic blood        pressure [BP]>180 mmHg or diastolic BP >100 mmHg) at screening        are excluded. Subjects requiring 2 or more medications to        control hypertension are eligible with Medical Monitor approval.    -   13. Subjects with known unstable or uncontrolled angina        pectoris.    -   14. Subjects with a known history of severe and/or uncontrolled        ventricular arrhythmias.    -   15. Subjects with a QTcF (QT corrected based on Fridericia's        equation) interval ≥450 msec or other factors that increase the        risk of QT prolongation or arrhythmic events (e.g., heart        failure, hypokalemia, family history of long QT interval        syndrome) at screening. Subjects with bundle branch block and a        prolonged QTc interval should be reviewed by the Medical Monitor        for potential inclusion.    -   16. Subjects taking medications that are known to prolong the QT        interval unless they can be transferred to other medications        within ≥5 half-lives prior to dosing.    -   17. Subjects with known infection with human immunodeficiency        virus (HIV) or active hepatitis B or C.    -   18. Subjects with any other medical or psychological condition,        deemed by the Investigator to be likely to interfere with a        subject's ability to sign informed consent, cooperate, or        participate in the study.    -   19. Subjects with known dysphagia, short-gut syndrome,        gastroparesis, or other conditions that limit the ingestion or        gastrointestinal absorption of drugs administered orally.    -   20. Subjects with clinical symptoms suggesting active central        nervous system (CNS) leukemia or known CNS leukemia. Evaluation        of cerebrospinal fluid is only required if there is a clinical        suspicion of CNS involvement by leukemia during screening.    -   21. Subjects with immediately life-threatening, severe        complications of leukemia such as uncontrolled bleeding,        pneumonia with hypoxia or shock, and/or disseminated        intravascular coagulation.    -   22. In the Phase 2 portion of the trial only, subjects who have        previously received treatment with an inhibitor of IDH.

Investigational Product, Dosage and Mode of Administration:

COMPOUND 1 (mesylate salt of2-methyl-1-[(4-[6-(trifluoromethyl)pyridin-2-yl]-6-{[2-(trifluoromethyl)pyridin-4-yl]amino}-1,3,5-triazin-2-yl)amino]propan-2-ol)will be provided as 5, 10, 25, 50, 100, 150 and 200 mg free-baseequivalent strength tablets to be administered orally.

Phase 1/Dose Escalation

The first 3 subjects in each cohort in the dose escalation portion ofthe study and the first 15 subjects in each arm of Part 1 Expansion willreceive a single dose of study drug on Day −3; their next dose of studydrug will be administered on C1D1 at which time subjects will startdaily dosing on Days 1 to 28 in 28-day cycles. Starting with C1D1,dosing is continuous; there are no inter-cycle rest periods. Subjectswho are not required to undergo the Day −3 PK/PD assessments willinitiate daily dosing with COMPOUND 1 on C1D1.

Subjects are required to fast (water is allowed) for 2 hours prior tostudy drug administration and for 1 hour following study drugadministration.

The dose of COMPOUND 1 administered to a subject will be dependent uponwhich dose cohort is open for enrollment when the subject qualifies forthe study. The starting dose of COMPOUND 1 to be administered to thefirst cohort of subjects is 30 mg administered orally twice a day, andthe maximum administered dose of COMPOUND 1 to be administered is 650 mgadministered orally once a day.

Phase 1/Part 1 Expansion and Phase 2

The starting dose of COMPOUND 1 recommended for evaluation is 100 mg QD.This is based on the safety, PK, pharmacodynamics and clinical activityof COMPOUND 1 observed to date in AG221-C-001. Evaluation ofpharmacodynamic response demonstrated sustained reduction in 2-HG plasmalevels by Day 1 of Cycle 2 and up to 98% inhibition in most subjectswith R140Q mutation at all doses. Increasing dose is associated withhigher exposure and inhibition of 2-HG in subjects with R172K mutation.Importantly, preliminary efficacy data of the 44 subjects treated at 100mg QD has shown an overall response rate of 36.4%. Thus a dose of 100 mgshould adequately achieve inhibition of 2-HG in subjects with eitherR140Q or R172K mutation. Moreover, the safety profile at 100 mg,including ≥Grade 3, is consistent with that of lower doses.

Intra-subject dose escalation is possible.

A subset of the clinical samples from the trial described in Example 1were analyzed at screening. Sample types included bone marrow,peripheral blood and mononuclear cells isolated from bone marrow orperipheral blood. DNAs were extracted from these samples and sequencedat Foundation Medicine (Heme Panel, seehttp://foundationone.com/learn.php) using next generation sequencingtechnique.

Duration of Treatment:

Subjects may continue treatment with COMPOUND 1 until diseaseprogression or development of unacceptable toxicity. Subjects whoexperience disease progression per the applicable response criteria whoare, in the opinion of the Investigator, benefiting from treatment maybe allowed to continue on study drug with approval of the MedicalMonitor.

End of Study:

End of study is defined as the time at which:

-   -   all subjects have discontinued treatment with COMPOUND 1 and        have been followed for survival for at least 12 months, or have        died, been lost to follow up, or withdrew consent prior to at        least 12 months of follow-up    -   or the date of receipt of the last data point from the last        subject that is required for primary, secondary and/or        exploratory analysis, as pre-specified in the protocol and/or        the Statistical Analysis Plan (SAP), whichever is the later        date.

Criteria for Evaluation:

Safety:

Monitoring of AEs, including determination of DLTs, serious adverseevents (SAEs), and AEs leading to discontinuation; safety laboratoryparameters; physical examination findings; vital signs; 12-lead ECGs;LVEF; and ECOG PS.

The severity of AEs will be assessed by the NCI CTCAE, Version 4.03.

Pharmacokinetics and Pharmacodynamics:

Serial blood sampling for determination of concentration-time profilesof COMPOUND 1 and its metabolite(6-(6-(trifluoromethyl)pyridin-2-yl)-N2-(2-(trifluoromethyl)pyridin-4-yl)-1,3,5-triazine-2,4-diamine).Urine sampling for determination of concentrations of COMPOUND 1 and itsmetabolite(6-(6-(trifluoromethyl)pyridin-2-yl)-N2-(2-(trifluoromethyl)pyridin-4-yl)-1,3,5-triazine-2,4-diamine)(dose escalation and Part 1 expansion subjects only). Blood and bonemarrow sampling for determination of 2-HG and α-KG levels.

Clinical Activity:

Serial blood and bone marrow sampling to determine response to treatmentbased on modified IWG response criteria or other appropriate responsecriteria based on the malignancy under study.

Overall response rate (ORR), the primary efficacy endpoint, is definedas the rate of responders including complete remission (CR), CR withincomplete platelet recovery (CRp), marrow CR (mCR) (morphologicleukemia-free state [MLFS] for subjects with AML), CR with incompletehematologic recovery (CRi), and partial remission (PR). Other measuresof clinical activity including complete remission rate (CRR), durationof remission/response, event-free survival, overall survival, and timeto remission/response will be summarized.

For Phase 1 Dose Escalation/Part 1 Expansion, the efficacy analysis ofresponse rates as assessed by the site Investigators using modifiedInternational Working Group (IWG) response criteria will be conducted inFull Analysis Set for each dose level, expansion arm, and overall ifappropriate. The analysis of Part 1 expansion arms may also includesubjects from the dose-escalation phase who received the samedose/regimen as subjects in the expansion arms and who meet theeligibility criteria of individual arms.

For Phase 2 portion of the trial, the primary efficacy analysis ofCOMPOUND 1 will be determined by the Investigators based on modifiedInternational Working Group (IWG) response criteria. Response will bealso be assessed retrospectively by an Independent Response AdjudicationCommittee (IRAC) using the Full Analysis Set (FAS). Key supportiveanalyses will be based on independent central review of response in FAS.

Analysis of a subset of the clinical samples from the trial for COMPOUND1 was conducted. FIG. 13 provides a visual of FLT mutations according toresponse categories for COMPOUND 1. FIG. 13 profiles bone marrow atscreening visit for patients only where genes were mutated in ≥2. InFIG. 13, genes (y-axis) are shown in decreasing order of frequency, withthe exception of IDH2, while patients (x-axis) are grouped by responsethen by similarity in alterations; only patients from dose escalationphase with an evaluable response included.

In certain embodiments, the patients with AML characterized by somaticmutations in FLT3 are resistant to treatment of COMPOUND 1.

In certain embodiments, a combination therapy with COMPOUND 1 and one ormore compounds that target a FLT3 pathway (e.g. quizartinib (AC220),sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412),lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028,ponatinib (AP24534), ASP2215, KW-2449, famitinib or DCC-2036) iseffective in treating AML in patients with AML characterized by somaticmutations in FLT3.

Example 2: A Phase 1, Multicenter, Open-Label, Dose-Escalation, Safety,Pharmacokinetic, Pharmacodynamic, and Clinical Activity Study of OrallyAdministered COMPOUND 2 in Subjects with Advanced HematologicMalignancies with an IDH1 Mutation

Indication: Treatment of patients with advanced hematologic malignancieswith an IDH1 mutation.

Objectives:

Primary:

To assess the safety and tolerability of treatment with COMPOUND 2administered continuously as a single agent dosed orally on Days 1 to 28of a 28-day cycle in subjects with advanced hematologic malignancies.The initial dosing regimen will be twice daily (approximately every 12hours). If warranted based on the emerging data, an alternative dosingschedule (e.g., once daily or three times daily), includingadministration of the same total daily dose using different dosingschedules in concurrent cohorts, may be explored as agreed upon by theClinical Study Team.

To determine the maximum tolerated dose (MTD) and/or the recommendedPhase 2 dose of COMPOUND 2 in subjects with advanced hematologicmalignancies.

To assess the clinical activity of AG-120 in subjects with relapsed orrefractory acute myelogenous leukemia (AML) with an IDH1 mutation whoare enrolled in Arm 1 of the expansion phase.

Secondary:

To describe the dose-limiting toxicities (DLTs) of COMPOUND 2 insubjects with advanced hematologic malignancies.

To characterize the pharmacokinetics (PK) of COMPOUND 2 in subjects withadvanced hematologic malignancies.

To evaluate the PK/pharmacodynamic (PD) relationship of COMPOUND 2 and2-hydroxygluturate (2 HG).

To characterize the clinical activity associated with COMPOUND 2 insubjects with advanced hematologic malignancies.

Methodology:

This study is a Phase 1, multicenter, open-label, dose-escalation,safety, PK/PD, and clinical activity evaluation of orally administeredCOMPOUND 2 in subjects with advanced hematologic malignancies thatharbor an IDH1 mutation. The study includes a dose escalation phase todetermine MTD and/or RP2D followed by expansion arms to further evaluatethe safety, tolerability, and clinical activity of COMPOUND 2 in selectpopulations.

Dose Escalation Phase

The dose escalation phase will utilize a standard “3+3” design. Duringthe dose escalation phase, consented eligible subjects with relapsed orrefractory AML, untreated AML ≥60 years of age who are not candidatesfor standard therapy, or myelodysplastic syndrome (MDS) with refractoryanemia with excess blasts will be enrolled into sequential cohorts ofincreasing doses of COMPOUND 2. Each dose cohort will plan to enroll aminimum of 3 subjects. The first 3 subjects enrolled in each dosingcohort during the dose escalation phase of the study will initiallyreceive a single dose of study drug on Day 3 (i.e., 3 days prior to thestart of daily dosing) and undergo PK/PD assessments over 72 hours toevaluate drug concentrations and 2-HG levels. The next dose of studydrug will be on Cycle 1 Day 1 (C1D1) at which time daily dosing willbegin. The initial dosing regimen was twice daily (approximately every12 hours). Based on the emerging data, a once daily (approximately every24 hours) dosing schedule has been implemented. Alternative dosingschedules, including administration of the same total daily dose usingdifferent dosing schedules in concurrent cohorts, may be explored asagreed upon by the Clinical Study Team. If there are multiple subjectsin the screening process at the time the third subject within a cohortbegins treatment, up to 2 additional subjects may be enrolled withapproval of the Medical Monitor. For these additional subjects, the Day3 through Day 1 PK/PD assessments may be considered optional followingdiscussion with the Medical Monitor.

The safety of dosing during the dose escalation phase will be evaluatedby the Clinical Study Team, comprised of the Sponsor (ResponsibleMedical Officer), Study Medical Monitor, and Investigators. The ClinicalStudy Team will review the emerging safety data from each cohort todetermine if dose escalation will occur.

Toxicity severity will be graded according to the National CancerInstitute Common Terminology Criteria for Adverse Events (NCI CTCAE)version 4.03. A DLT is defined as outlined below.

Non-Hematologic:

All non-hematologic toxicities CTCAE≥Grade 3.

Hematologic:

Prolonged myelosuppression with persistence of ≥Grade 4 neutropenia orthrombocytopenia in the absence of leukemia (blast count <5%) at least42 days after the initiation of Cycle 1 therapy.

Due to frequent co-morbidities and concurrent medications in thepopulation under study, attribution of adverse events (AEs) to aparticular drug is challenging. Therefore, all AEs that cannot clearlybe determined to be unrelated to COMPOUND 2 will be considered relevantto determining DLTs and will be reviewed by the Clinical Study Team. TheClinical Study Team also will review any other emergent toxicities thatare not explicitly defined by the DLT criteria to determine if anywarrant a DLT designation.

If, after the third subject completes the 28-day DLT evaluation period(i.e., Cycle 1), no DLTs are observed, the study will proceed with doseescalation to the next cohort following safety review by the ClinicalStudy Team. If 1 of 3 subjects experiences a DLT during the first cycle,3 additional subjects will be enrolled in that cohort. If none of theadditional 3 subjects experience a DLT, dose escalation may continue tothe next cohort following safety review by the Clinical Study Team. If 2or more subjects in a cohort experience DLTs during the first cycle,dose escalation will be halted and the next lower dose level will bedeclared the MTD. Alternatively, a dose level intermediate between thedose level exceeding MTD and the previous does level may be explored anddeclared MTD if <2 out of 6 subjects experience a DLT at that dose. Ifthe MTD cohort included only 3 subjects, an additional 3 subjects willbe enrolled at that dose level to confirm that <2 of 6 subjectsexperience a DLT at that dose.

Increases in the dose of AG-120 for each dose cohort will be guided byan accelerated titration design, where the daily dose may be doubled(100% increase) from one cohort to the next until AG-120-related CTCAEGrade 2 or greater toxicity is observed in any subject within thecohort. Following evaluation by the Clinical Study Team, subsequentincreases in dose will be guided by the observed toxicity, andpotentially PK and PK/PD data, until the MTD is determined. The absolutepercent increase in the daily dose will be determined by the ClinicalStudy Team predicated on the type and severity of any toxicity seen inthe prior dose cohorts (but will never exceed 100%). The MTD is thehighest dose that causes DLTs in <2 of 6 subjects.

If no DLTs are identified during the dose escalation phase, doseescalation may continue for at least 2 dose levels above the projectedmaximum clinically effective exposure, as determined by an ongoingassessment of PK/PD and any observed clinical activity; this may occurin parallel with the expansion phase.

To optimize the number of subjects treated at a potentially clinicallyrelevant dose, intra-subject dose escalation will be permitted withapproval of the Medical Monitor.

Expansion Phase

Following determination of the recommended dose and dosing regimen fromthe dose escalation phase, the expansion phase will open to furtherexplore the dose in subjects with specific hematologic malignancies.During the expansion phase, 4 non-randomized arms of approximately 25subjects per arm with IDH1-mutated hematologic malignancies will beenrolled as follows:

Arm 1: Relapsed or refractory AML defined as:

-   -   Subjects who relapse after transplantation;    -   Subjects in second or later relapse;    -   Subjects who are refractory to initial induction or reinduction        treatment.    -   Subjects who relapse within 1 year of initial treatment,        excluding patients with favorable-risk status according to NCCN        Guidelines, version 1.2015.

Arm 2: Untreated AML who are not candidates for standard therapy due toage, comorbid condition, performance status, and/or adverse riskfactors, according to the Investigator and with approval of the MedicalMonitor.

Arm 3: Other non-AML IDH1-mutated relapsed and/or refractory advancedhematologic malignancies, where no standard of care treatment option isavailable. Such as:

-   -   Myelodysplastic syndrome that is recurrent or refractory after        having failed hypomethylating agent(s) and with the approval of        Medical Monitor.    -   Relapsed and/or primary refractory chronic myelomonocytic        leukemia [CMML] with the approval of Medical Monitor.    -   Other non-AML IDH1-mutated relapsed and/or refractory advanced        hematologic malignancy, that have failed standard of care or no        standard of care treatment option is available according to the        Investigator and with the approval of the Medical Monitor.

Arm 4: Relapsed AML patients not eligible for Arm 1 that have failedavailable standard of care or are unable to receive standard of care dueto age, comorbid condition, performance status, and/or adverse riskfactors, according to the Investigator and with approval of the MedicalMonitor.

An interim analysis for safety and futility will be conducted for Arm 1of the expansion phase at the time at which the first 25 subjects havebeen treated and followed for 2 cycles or discontinued earlier. If theobjective response rate (ORR), defined as including all responses ofcomplete remission (CR), CR with incomplete platelet recovery (CRp),morphologic leukemia-free state (MLFS), CR with incomplete hematologicrecovery (CRi), and partial remission (PR), is <15% (i.e., <4responders), then additional enrollment into Arm 1 may be terminated; ifthe ORR is ≥15%, an additional approximately 100 subjects will beenrolled. Enrollment will not be held for the interim analysis.

General Study Conduct

Following informed consent, all subjects will undergo screeningprocedures within 28 days prior to C1D1 to determine eligibility. Allsubjects are required to have confirmation of IDH1 R132 gene-mutateddisease from a bone marrow aspirate and/or biopsy. For subjects in thedose escalation phase, documentation can be based on local site testingwith central laboratory testing performed retrospectively. Subjects inthe expansion phase are required to have IDH1 R132 gene-mutated diseasebased on central laboratory testing during screening prior to treatment.Additional screening procedures include medical, surgical, andmedication history; a buccal swab for germ-line mutation analysis;complete physical examination; vital signs; Eastern Cooperative OncologyGroup (ECOG) performance status (PS); 12-lead electrocardiogram (ECG);left ventricular ejection fraction (LVEF); clinical laboratoryassessments (hematology, chemistry, coagulation, urinalysis, and serumpregnancy test); bone marrow biopsy and/or aspirate; and blood and bonemarrow samples for 2-HG measurement and other exploratory assessments.In addition, subjects in the dose escalation phase will have urinesamples for 2 HG measurement and blood samples for determination ofplasma cholesterol and 4β-OH-cholesterol levels during screening.

Dose Escalation Phase:

Three days prior to starting the daily dosing of COMPOUND 2 (Day −3),the first 3 subjects enrolled in each cohort in the dose escalationphase will receive a single dose of COMPOUND 2 in clinic and have serialblood and urine samples obtained for determination of concentrations ofCOMPOUND 2 and 2-HG. A full 72-hour PK/PD profile will be conducted:subjects will be required to remain at the study site for 10 hours onDay −3 and return on Days −2, −1, and 1 for 24, 48, and 72 hour samples,respectively. Daily treatment with COMPOUND 2 will begin on C1D1;subjects in the dose escalation phase who did not undergo the Day −3PK/PD assessments are to remain in clinic for 4 hours after the C1D1dose for clinical observation.

Subjects in the dose escalation phase also will undergo PK/PDassessments over a 10-hour period on both C1D15 and C2D1. Additionalpre-dose urine and/or blood sampling will be conducted on C1D8, C1D22,C2D15, C3D1, C3D15, and on Day 1 of all subsequent cycles fordetermination of COMPOUND 2 and 2-HG concentration. Available bonemarrow biopsy samples also will be assessed for 2-HG levels.

Expansion Phase: Subjects in the expansion phase are not required toundergo the Day −3 assessments; these subjects will undergo an 8-hourPK/PD profile conducted on Day 1 of Cycles 1 and 2. Additional bloodsamples for PK/PD assessments will be drawn pre-dose (within 30 minutes)on Days 8 and 15 of Cycle 1, Day 1 of Cycle 3, on Day 1 of allsubsequent cycles, and at the End of Treatment visit. Time-matched12-lead ECGs will be conducted in triplicate on Day 1 of Cycles 1 and 2;a triplicate ECG also will be conducted at the End of Treatment visit.Single 12-lead ECGs will be conducted at Screening, 4 hours post-dose onDays 8 and 15 of Cycle 1, on Day 1 of every cycle beginning with Cycle3, and at the Follow-up visit. Available bone marrow biopsy samples willbe assessed for 2-HG levels.

Other Safety Assessments:

All subjects will undergo safety assessments during the treatment periodto include physical examination, vital signs, ECOG PS, 12-lead ECGs,LVEF, and clinical laboratory assessments (hematology, chemistry,coagulation, urinalysis, and pregnancy testing).

Clinical Activity Assessments:

All subjects will have the extent of their disease assessed, includingbone marrow biopsies and/or aspirates and peripheral blood, atscreening, on Day 15 (dose escalation phase only), on Day 29, every 28days thereafter through Month 12, and then every 56 days thereafterwhile on study drug treatment, independent of dose delays and/or doseinterruptions, and/or at any time when progression of disease issuspected. Note that the Day 15 bone marrow evaluation during doseescalation should not be used to determine study treatment continuationstatus. Response to treatment and treatment decisions in all subjectswill be determined by the Investigators based on modified InternationalWorking Group (IWG) response criteria or other appropriate responsecriteria for the malignancy under study. For subjects with relapsed orrefractory AML enrolled in the expansion phase, response also will beassessed by an Independent Review Committee.

End of Treatment and Follow-up:

Subjects may continue treatment with COMPOUND 2 until diseaseprogression, development of other unacceptable toxicity, confirmedpregnancy, undergoing a hematopoietic stem cell transplant (HSCT),death, withdrawal of consent, lost to follow-up, or Sponsor ending thestudy, whichever occurs first. Subjects who experience diseaseprogression per the applicable response criteria who are, in the opinionof the Investigator, benefiting from treatment may be allowed tocontinue on study drug with approval of the Medical Monitor.

All subjects are to undergo an end of treatment assessment (withinapproximately 5 days of the last dose of study drug); in addition, afollow-up safety assessment is to be scheduled 28 days after the lastdose.

Subjects who achieve an adequate response to treatment with COMPOUND 2and meet other criteria required to undergo HSCT may proceed to HSCTafter discontinuation of COMPOUND 2 and will be followed on study fordisease evaluation (approximately monthly, as standard of care) and anynew bone marrow transplant (BMT) conditioning antineoplastic therapiesreceived until disease relapse, death, withdrawal of consent, lost tofollow-up, or end of study. If a subject discontinues COMPOUND 2 toundergo HSCT, but is then deemed ineligible for HSCT, the subject mayrestart COMPOUND 2 with Medical Monitor approval. Subjects who fail HSCTand have recurrent IDH1-mutant positive disease may be eligible torestart treatment with COMPOUND 2 with Medical Monitor approval.

All subjects, including those who relapse following HSCT and elect notto restart treatment, will enter survival follow-up and will becontacted monthly for assessment of survival status and BMT conditioningantineoplastic therapies since discontinuation of study drug untildeath, withdrawal of consent, lost to follow-up, or end of study.

Concomitant medications do not need to be avoided while subjects are nolonger receiving COMPOUND 2 ie, HSCT or survival follow-up periods).

Number of Subjects (Planned):

It is estimated that approximately 236 subjects will be enrolled in thestudy.

Assuming that identification of the MTD requires the evaluation of 7dose levels of AG-120 with 3 to 5 subjects per dose level, with theexception that the MTD requires 6 subjects, then 36 subjects will beenrolled during the dose escalation part of the study.

Four cohorts of approximately 25 subjects each in specific hematologicmalignancies (total 100 subjects) will initially be enrolled in theexpansion phase of the study with the possible additional enrollment of100 subjects with relapsed or refractory AML in Arm 1 depending onreview of safety and clinical activity at the interim analysis.

Additional subjects may be enrolled during dose escalation, for thereplacement of subjects who are not evaluable for the assessment of doseescalation, for evaluation of alternative dosing regimens or for furtherexploring safety, PK, PK/PD, or preliminary clinical activity used toguide the selection of the RP2D.

Diagnosis and Main Criteria for Inclusion:

Inclusion Criteria:

-   -   Subjects must meet all of the following criteria to be enrolled        in the study:    -   Subjects must be ≥18 years of age.    -   Subjects must have an advanced hematologic malignancy including:

Dose Escalation Phase:

Relapsed and/or primary refractory AML as defined by World HealthOrganization (WHO) criteria; or

Untreated AML, ≥60 years of age and are not candidates for standardtherapy due to age, performance status, and/or adverse risk factors,according to the treating physician and with approval of the MedicalMonitor;

Myelodysplastic syndrome with refractory anemia with excess blasts(subtype RAEB-1 or RAEB-2), or considered high-risk by the RevisedInternational Prognostic Scoring System (IPSS-R) Greenberg et al. Blood.2012; 120(12):2454-65 that is recurrent or refractory, or the subject isintolerant to established therapy known to provide clinical benefit fortheir condition (i.e., subjects must not be candidates for regimensknown to provide clinical benefit), according to the treating physicianand with approval of the Medical Monitor.

(Subjects with other relapsed and/or primary refractory hematologiccancers, for example CMML, who fulfill the inclusion/excluding criteriamay be considered on a case-by case basis, with approval of the MedicalMonitor.)

Expansion Phase:

Arm 1: Relapsed or refractory AML, defined as:

-   -   Subjects who relapse after transplantation;    -   Subjects in second or later relapse;    -   Subjects who are refractory to initial induction or reinduction        treatment.    -   Subjects who relapse within 1 year of initial treatment,        excluding patients with favorable-risk status according to NCCN        Guidelines, version 1.2015.

Arm 2: Untreated AML who are not candidates for standard therapy due toage, cormorbid condition, performance status, and/or adverse riskfactors, according to the Investigator and with approval of the MedicalMonitor.

Arm 3: Other non-AML IDH1-mutated relapsed and/or refractory advancedhematologic malignancies, where no standard of care treatment option isavailable. Such as:

-   -   Myelodysplastic syndrome that is recurrent or refractory after        having failed hypomethylating agent(s) and with the approval of        Medical Monitor.    -   Relapsed and/or primary refractory CMML with the approval of        Medical Monitor    -   Other non-AML IDH1-mutated relapsed and/or refractory advanced        hematologic malignancy, that have failed standard of care or no        standard of care treatment option is available according to the        Investigator and with the approval of the Medical Monitor.

Arm 4: Relapsed AML patients not eligible for Arm 1 that have failedavailable standard of care or are unable to receive standard of care dueto age, comorbid condition, performance status, and/or adverse riskfactors, according to the Investigator and with approval of the MedicalMonitor.

Subjects must have documented IDH1 R132 gene-mutated disease

-   -   For subjects in the dose escalation phase, IDH1 mutation may be        based on local evaluation. (Centralized testing will be        performed retrospectively.)    -   For subjects in the expansion phase, central testing of IDH1        gene-mutated disease is required during screening to confirm        eligibility.

Subjects must be amenable to serial bone marrow sampling, peripheralblood sampling, and urine sampling during the study.

-   -   The diagnosis and evaluation of AML or MDS will be made by bone        marrow aspiration and/or biopsy. If an aspirate is unobtainable        (i.e., a “dry tap”), the diagnosis may be made from the core        biopsy.

Subject must be able to understand and willing to sign an informedconsent. A legally authorized representative may consent on behalf of asubject who is otherwise unable to provide informed consent, ifacceptable to and approved by the site and/or site's InstitutionalReview Board (IRB).

Subjects must have ECOG PS of 0 to 2.

Platelet count ≥20,000/μL (Transfusions to achieve this level areallowed.) Subjects with a baseline platelet count of <20,000/μL due tounderlying malignancy are eligible with Medical Monitor approval.

Subjects must have adequate hepatic function as evidenced by:

-   -   Serum total bilirubin ≤1.5× upper limit of normal (ULN), unless        considered due to Gilbert's disease or leukemic involvement;    -   Aspartate aminotransferase (AST), alanine aminotransferase        (ALT), and alkaline phosphatase (ALP)≤3.0×ULN, unless considered        due to leukemic involvement.

Subjects must have adequate renal function as evidenced by:

-   -   Serum creatinine ≤2.0×UL OR    -   Creatinine clearance ≥40 mL/min based on the Cockroft-Gault        glomerular filtration rate (GFR) estimation: (140−Age)×(weight        in kg)×(0.85 if female)/72×serum creatinine

Subjects must be recovered from any clinically relevant toxic effects ofany prior surgery, radiotherapy, or other therapy intended for thetreatment of cancer. (Subjects with residual Grade 1 toxicity, forexample Grade 1 peripheral neuropathy or residual alopecia, are allowedwith approval of the Medical Monitor.)

Female subjects with reproductive potential must agree to undergomedically supervised pregnancy test prior to starting study drug. Thefirst pregnancy test will be performed at screening (within 7 days priorto first study drug administration), and on the day of the first studydrug administration and confirmed negative prior to dosing and Day 1before dosing all subsequent cycles.

Female subjects with reproductive potential must have a negative serumpregnancy test within 7 days prior to the start of therapy. Subjectswith reproductive potential are defined as sexually mature women whohave not undergone a hysterectomy, bilateral oophorectomy or tubalocclusion or who have not been naturally postmenopausal (i.e., who havenot menstruated at all) for at least 24 consecutive months (i.e., hashad menses at any time in the preceding 24 consecutive months). Femalesof reproductive potential as well as fertile men and their partners whoare female of reproductive potential must agree to abstain from sexualintercourse or to use two highly effective forms of contraception fromthe time of giving informed consent, during the study and for 90 days(females and males) following the last dose of COMPOUND 2. A highlyeffective form of contraception is defined as hormonal oralcontraceptives, injectables, patches, intrauterine devices,double-barrier method (e.g., synthetic condoms, diaphragm, or cervicalcap with spermicidal foam, cream, or gel), or male partnersterilization.

Exclusion Criteria:

Subjects who meet any of the following criteria will not be enrolled inthe study:

Subjects who previously received prior treatment with a mutant-specificIDH1 inhibitor and progressed on therapy.

Subjects who have undergone hematopoietic stem cell transplant (HSCT)within 60 days of the first dose of COMPOUND 2, or subjects onimmunosuppressive therapy post HSCT at the time of screening, or withclinically significant graft-versus-host disease (GVHD). (The use of astable dose of oral steroids post HSCT and/or topical steroids forongoing skin GVHD is permitted with Medical Monitor approval.)

Subjects who received systemic anticancer therapy or radiotherapy <14days prior to their first day of study drug administration. (Hydroxyureais allowed prior to enrollment and after the start of COMPOUND 2 for thecontrol of peripheral leukemic blasts in subjects with leukocytosis(white blood cell [WBC] counts >30,000/μL).

Subjects who received an investigational agent <14 days prior to theirfirst day of study drug administration. In addition, the first dose ofCOMPOUND 2 should not occur before a period ≥5 half-lives of theinvestigational agent has elapsed.

Subjects taking sensitive cytochrome P450 (CYP) 3A4 substratemedications are excluded from the study unless they can be transferredto other medications within ≥5 half-lives prior to dosing, or unless themedications can be properly monitored during the study.

Subjects taking P-glycoprotein (P-gp) transporter-sensitive substratemedications are excluded from the study unless they can be transferredto other medications within ≥5 half-lives prior to dosing, or unless themedications can be properly monitored during the study.

Subjects for whom potentially curative anticancer therapy is available.

Subjects who are pregnant or breast feeding.

Subjects with an active severe infection that required anti-infectivetherapy or with an unexplained fever >38.5° C. during screening visitsor on their first day of study drug administration (at the discretion ofthe Investigator, subjects with tumor fever may be enrolled).

Subjects with known hypersensitivity to any of the components ofCOMPOUND 2.

Subjects with New York Heart Association (NYHA) Class III or IVcongestive heart failure or LVEF<40% by echocardiogram (ECHO) ormulti-gated acquisition (MUGA) scan obtained within approximately 28days of C1D1.

Subjects with a history of myocardial infarction within the last 6months of screening.

Subjects with known unstable or uncontrolled angina pectoris.

Subjects with a known history of severe and/or uncontrolled ventriculararrhythmias.

Subjects with QTc interval ≥450 msec or with other factors that increasethe risk of QT prolongation or arrhythmic events (e.g., heart failure,hypokalemia, family history of long QT interval syndrome) at screening.Subjects with bundle branch block and a prolonged QTc interval should bereviewed by the Medical Monitor for potential inclusion.

Subjects taking medications that are known to prolong the QT intervalunless they can be transferred to other medications within ≥5 half-livesprior to dosing or unless the medications can be properly monitoredduring the study.

Subjects with known infection with human immunodeficiency virus (HIV) oractive hepatitis B or C.

Subjects with any other medical or psychological condition, deemed bythe Investigator to be likely to interfere with a subject's ability tosign informed consent, cooperate, or participate in the study.

Subjects with known dysphagia, short-gut syndrome, gastroparesis, orother conditions that limit the ingestion or gastrointestinal absorptionof drugs administered orally.

Subjects with clinical symptoms suggesting active central nervous system(CNS) leukemia or known CNS leukemia. Evaluation of cerebrospinal fluidis only required if there is a clinical suspicion of CNS involvement byleukemia during screening.

Subjects with immediately life-threatening, severe complications ofleukemia such as uncontrolled bleeding, pneumonia with hypoxia or shock,and/or disseminated intravascular coagulation.

Investigational Product, Dosage and Mode of Administration

COMPOUND 2 will be provided as 10, 50, 200, and 250 mg strength tabletsto be administered orally.

The first 3 subjects in each dose escalation cohort in the doseescalation portion of the study will receive a single dose of study drugon Day −3; their next dose of study drug will be administered on C1D1 atwhich time subjects will start dosing daily on Days 1 to 28 in 28-daycycles, with plans to explore alternative dosing regimens if warranted.Starting with C1D1, dosing is continuous; there are no inter-cycle restperiods. Subjects who are not required to undergo the Day −3 PK/PDassessments will initiate daily dosing with COMPOUND 2 on C1D1.

The dose of COMPOUND 2 administered to a subject will be dependent uponwhich dose cohort is open for enrollment when the subject qualifies forthe study. The starting dose of AG-120 to be administered to the firstcohort of subjects in the dose escalation phase is 100 mg administeredorally twice a day (200 mg/day). The starting dose and regimen forsubjects in the expansion phase (500 mg QD) was based on safety, PK,PK/PD, and clinical activity results from the dose-escalation phase ofthe study will be provided as 10, 50 and 200 mg strength tablets to beadministered orally.

Duration of Treatment:

Subjects may continue treatment with COMPOUND 2 until diseaseprogression, development of other unacceptable toxicity, confirmedpregnancy, undergoing HSCT, death, withdrawal of consent, lost tofollow-up, or Sponsor ending the study, whichever occurs first. Subjectswho experience disease progression per the applicable response criteriawho are, in the opinion of the Investigator, benefiting from treatmentmay be allowed to continue on study drug with approval of the MedicalMonitor.

End of Study:

End of study is defined as the time at which all subjects havediscontinued treatment with COMPOUND 2 and have been followed forsurvival for at least 12 months, or have died, been lost to follow up,or withdrew consent prior to 12 months of follow-up.

Criteria for Evaluation:

Safety:

Monitoring of AEs, including determination of DLTs, serious adverseevents (SAEs), and AEs leading to discontinuation; safety laboratoryparameters; physical examination findings; vital signs; 12-lead ECGs;LVEF; and ECOG PS.

The severity of AEs will be assessed by the NCI CTCAE, version 4.03.

Pharmacokinetics and Pharmacodynamics:

Serial blood sampling for determination of concentration-time profilesof COMPOUND 2. Blood and bone marrow sampling for determination of 2-HGlevels. Urine sampling for determination of urinary concentrations ofCOMPOUND 2 and 2-HG levels (dose escalation subjects only).

Clinical Activity:

Serial blood and bone marrow sampling to determine response to treatmentbased on modified IWG response criteria in AML or other appropriateresponse criteria based on the malignancy under study.

Response to treatment as assessed by the site Investigators usingmodified IWG or other appropriate response criteria for the diseaseunder study will be tabulated. Response will be summarized by bestobjective response categories, complete remission rate (CRR), andobjective response rate (ORR), including all responses of CR, CRp, mCR(morphologic leukemia-free state [MLFS] for subjects with AML), CRi, andPR. Other measures of clinical activity, including duration of completeremission, duration of response, event-free survival, overall survival,and time to remission/response will be summarized.

The primary analysis of the clinical activity of COMPOUND 2 for theexpansion phase will be based on Investigator review of response (CRR,ORR, and duration of remission/response) using the Full Analysis Set.Key supportive analyses will be based on the central independent review.Additional efficacy analyses may be conducted using the EfficacyAnalysis Set.

A analysis of a subset of the clinical samples from the trial forCOMPOUND 2 was conducted. FIG. 14 provides a visual of FLT mutationsaccording to response categories for COMPOUND 2. FIG. 14 profiles bonemarrow at screening visit for patients only where genes were mutated in≥2. In FIG. 14, genes (y-axis) are shown in decreasing order offrequency, with the exception of IDH2, while patients (x-axis) aregrouped by response then by similarity in alterations; only patientsfrom dose escalation phase with an evaluable response included.

In certain embodiments, the patients with AML characterized by somaticmutations in FLT3 are resistant to treatment of COMPOUND 2.

In certain embodiments, a combination therapy with COMPOUND 2 and one ormore compounds that target a FLT3 pathway (e.g. FLT3 inhibitor selectedfrom quizartinib (AC220), sunitinib (SU11248), sorafenib (BAY 43-9006),midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596),PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449,famitinib or DCC-2036) is effective in treating AML in patients with AMLcharacterized by somatic mutations in FLT3.

Having thus described several aspects of several embodiments, it is tobe appreciated various alterations, modifications, and improvements willreadily occur to those skilled in the art. Such alterations,modifications, and improvements are intended to be part of thisdisclosure, and are intended to be within the spirit and scope of theinvention. Accordingly, the foregoing description and drawings are byway of example only.

1. A method of treating a hematological malignancy in a subjectcomprising administering to the subject a mutant isocitratedehydrogenase 1 (IDH1) inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide,having the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2),wherein the hematological malignancy is a malignancy characterized bythe presence of a mutant allele of IDH1 and the absence of a FLT3mutation.
 2. A method of treating a solid tumor in a subject comprisingadministering to the subject a mutant isocitrate dehydrogenase 1 (IDH1)inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide,having the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2),wherein the solid tumor is characterized by the presence of a mutantallele of IDH1 and the absence of a FLT3 mutation.
 3. A method oftreating a hematological malignancy in a subject comprisingadministering to the subject a mutant isocitrate dehydrogenase 1 (IDH1)inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide,having the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2)in combination with a FLT3 inhibitor, wherein the hematologicalmalignancy is a malignancy characterized by the presence of a mutantallele of IDH1 and a mutant FLT3.
 4. A method of treating a solid tumorin a subject comprising administering to the subject a mutant isocitratedehydrogenase 1 (IDH1) inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide,having the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2)in combination with a FLT3 inhibitor, wherein the solid tumorcharacterized by the presence of a mutant allele of IDH1 and a mutantFLT3.
 5. The method of any one of claims 1 to 4, wherein the IDH1mutation is an IDH1 R132X mutation.
 6. The method of claim 5, whereinthe IDH1 mutation is an IDH1 R132H, R132C, R132L, R132V, R132S or R132Gmutation.
 7. The method of any one of claims 3 to 6, wherein the FLT3inhibitor is selected from quizartinib (AC220), sunitinib (SU11248),sorafenib (BAY 43-9006), midostaurin (PKC412), lestaurtinib (CEP-701),crenolanib (CP-868596), PLX3397, E6201, AKN-028, ponatinib (AP24534),ASP2215, KW-2449, famitinib and DCC-2036.
 8. The method of claim 1 or 3,wherein the malignancy is acute myelogenous leukemia (AML),myelodysplastic syndrome (MDS, myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), each characterized by thepresence of a mutant allele of IDH1 and the method comprisesadministering a therapeutically effective amount of COMPOUND 2 to thesubject.
 9. The method of claim 8, wherein the malignancy is acutemyelogenous leukemia (AML), characterized by the presence of a mutantallele of IDH1.
 10. The method of claim 2 or 4, wherein the solid tumoris glioma, melanoma, chondrosarcoma, cholangiocarcinoma (includingintrahepatic cholangiocarcinoma (IHCC)), prostate cancer, colon cancer,or non-small cell lung cancer (NSCLC), each characterized by thepresence of a mutant allele of IDH1 and the method comprisesadministering a therapeutically effective amount of COMPOUND 2 to thesubject.
 11. The method of any one of claims 1 to 10, wherein the doseof COMPOUND 2 is administered at a dose of about 20 to about 2000mg/day.
 12. The method of any one of claims 1 to 10, wherein the dose ofCOMPOUND 2 is administered at a dose of about 50 to about 500 mg/day.13. A method of identifying a cancer subject suitable for treatment withan IDH1 inhibitor, comprising: (a) obtaining a biological sample from asubject having cancer; (b) screening the biological sample for an IDH1mutation and a FLT3 mutation; and (c) if the cancer is characterized bythe presence of a mutant allele of IDH1 and the absence of a FLT3mutation, identifying the subject as a cancer subject suitable fortreatment with an IDH1 inhibitor.
 14. The method of claim 13, whereinthe IDH1 inhibitor is COMPOUND
 2. 15. A method of identifying a cancersubject suitable for treatment with a combination of an IDH1 inhibitorand a FLT3 pathway inhibitor, comprising: (a) obtaining a biologicalsample from a subject having cancer; (b) screening the biological samplefor an IDH1 mutation and a FLT3 mutation; and (c) if the cancer ischaracterized by the presence of a mutant allele of IDH1 and a mutantFLT3, identifying the subject as a cancer subject suitable for treatmentwith a combination therapy with an IDH1 inhibitor and a FLT3 inhibitor.16. The method of claim 15, wherein the IDH1 inhibitor is COMPOUND 2.17. The method of claim 15 or 16, wherein the FLT3 inhibitor is selectedfrom quizartinib (AC220), sunitinib (SU11248), sorafenib (BAY 43-9006),midostaurin (PKC412), lestaurtinib (CEP-701), crenolanib (CP-868596),PLX3397, E6201, AKN-028, ponatinib (AP24534), ASP2215, KW-2449,famitinib and DCC-2036.
 18. The method of any one of claims 13 to 17,wherein the cancer is a solid tumor or a hematologic malignancy.
 19. Themethod of claim 18, wherein the hematologic malignancy is AML.
 20. Themethod of claim 19, wherein the AML is relapsed or refractory.
 21. Themethod of any one of claims 1 to 20, wherein the mutant FLT3 is FLT3-ITDor FLT3-KDM.
 22. The method of any one of claims 1 to 21, wherein thethe mutant FLT3 is FLT3-ITD.
 23. A compound for use in a method oftreating a hematological malignancy in a subject, wherein the compoundis a mutant isocitrate dehydrogenase 1 (IDH1) inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamidehaving the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2),wherein the hematological malignancy is a malignancy characterized bythe presence of a mutant allele of IDH1 and the absence of a FLT3mutation.
 24. A compound for use in a method of treating a solid tumorin a subject, wherein the compound is a mutant isocitrate dehydrogenase1 (IDH1) inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamidehaving the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2),wherein the solid tumor is characterized by the presence of a mutantallele of IDH1 and the absence of a FLT3 mutation.
 25. A compound foruse in a method of treating a hematological malignancy in a subject,wherein the compound is a mutant isocitrate dehydrogenase 1 (IDH1)inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamidehaving the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2)in combination with a FLT3 inhibitor, wherein the hematologicalmalignancy is a malignancy characterized by the presence of a mutantallele of IDH1 and a mutant FLT3.
 26. A compound for use in a method oftreating a solid tumor in a subject, wherein the compound is a mutantisocitrate dehydrogenase 1 (IDH1) inhibitor(S)—N—((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamidehaving the following formula:

or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer,isotopologue, prodrug, metabolite, or a polymorph thereof (COMPOUND 2)in combination with a FLT3 inhibitor, wherein the solid tumor ischaracterized by the presence of a mutant allele of IDH1 and a mutantFLT3.
 27. The compound for use of any one of claims 23 to 26, whereinthe IDH1 mutation is an IDH1 R132H, R132C, R132L, R132V, R132S or R132Gmutation.
 28. The compound for use of any one of claims 25 to 27,wherein the FLT3 inhibitor is selected from quizartinib (AC220),sunitinib (SU11248), sorafenib (BAY 43-9006), midostaurin (PKC412),lestaurtinib (CEP-701), crenolanib (CP-868596), PLX3397, E6201, AKN-028,ponatinib (AP24534), ASP2215, KW-2449, famitinib and DCC-2036.
 29. Thecompound for use of claim 23 or 25, wherein the malignancy is acutemyelogenous leukemia (AML), myelodysplastic syndrome (MDS,myeloproliferative neoplasms (MPN), chronic myelomonocytic leukemia(CMML), B-acute lymphoblastic leukemias (B-ALL), or lymphoma (e.g.,T-cell lymphoma), each characterized by the presence of a mutant alleleof IDH1 and the method comprises administering a therapeuticallyeffective amount of COMPOUND 2 to the subject.
 30. The compound for useof claim 29, wherein the malignancy is acute myelogenous leukemia (AML),characterized by the presence of a mutant allele of IDH1.
 31. Thecompound for use of claim 30, wherein the AML is relapsed or refractory.32. The compound for use of claim 24 or 26, wherein the solid tumor isglioma, melanoma, chondrosarcoma, cholangiocarcinoma (includingintrahepatic cholangiocarcinoma (IHCC), prostate cancer, colon cancer,or non-small cell lung cancer (NSCLC), each characterized by thepresence of a mutant allele of IDH1 and the method comprisesadministering a therapeutically effective amount of COMPOUND 2 to thesubject.
 33. The compound for use of any one of claims 23 to 32, whereinthe dose of COMPOUND 2 is about 20 to about 2000 mg/day or about 50 toabout 500 mg/day.
 34. The compound for use of any one of claims 25 to33, wherein the mutant FLT3 is FLT3-ITD or FLT3-KDM.
 35. The compoundfor use of any one of claims 25 to 34, wherein the mutant FLT3 isFLT3-ITD.